Introduction: Physical inactivity is thought to contribute to the emergence of non-communicable diseases in post-conflict settings of low-income countries. Sport-for-development (SfD) organisations in these regions claim to improve the health of programme participants. However, there is a paucity of supporting evidence. I assessed the impact of a voluntary community-based SfD intervention on the physical activity (PA), physical fitness (PF) and mental health (MH) of adolescents in Gulu, Uganda. Methods: The Acholi Psychosocial Assessment Instrument (APAI), standing broad jump (SBJ), multi-stage fitness test (MSFT) and BMI-for-age (BFA) were adapted to the local context. I tested their feasibility and reliability with a repeat-measures design (n=70). A cross-sectional analysis of a random sample was used to assess the local needs and establish the PF and MH of the adolescents reached by the intervention (n=1464). This was also the baseline assessment for the impact evaluation. It comprised a randomised control trial (n=144) nested within a cohort study (n=1400) and triangulated by cross-sectional assessment of PA using accelerometry (n=54). Results: The adapted PF and MH measures demonstrated good intra-tester reliability (ICC>0.75). Adolescents in Gulu predominantly had “healthy” BFA (>90%). They performed better than global norms for the SBJ (p<0.001), but worse for the MSFT (p<0.05). The girls who registered for the intervention had higher PF at baseline (p<0.05) and experienced no significant benefits when compared to the community. The aerobic capacity of the boys intervention group increased relative to the community (p<0.01), but was not significantly different to the trial control group whose PF also improved. The PA results concurred with this finding. Boys in the intervention group experienced a deterioration in MH relative to their peers (p<0.05). Implications: It is feasible to apply rigorous evaluation methods to SfD interventions. Although adolescents in Gulu have poor aerobic capacity, a voluntary programme may not reach those at risk. Interpreting the impact evaluation was limited by a lack of programme development theory, but suggested that opportunities for non-competitive play may confer PF benefit without harming MH. Further investigation is warranted.

The envelope glycoprotein (Env) of Human Immunodeficiency Virus 1 (HIV-1) has developed several immune-evasion mechanisms to avoid the induction of neutralising antibodies, including immunodominant non-neutralising epitopes, conformational flexibility of conserved epitopes, and spontaneous subunit dissociation, thus impeding vaccine development. Here, chemical modification of Env-based vaccine antigens is explored to overcome these obstacles. Firstly, covalent fixation of Env by chemical cross-linking was used to stabilise the conformationally flexible structure and prevent subunit dissociation. Cross-linked Env constructs showed reduced binding of many non-neutralising antibodies whilst largely maintaining antibody recognition by broadly neutralising antibodies. Compared to unmodified material, immunisation with some of these cross-linked proteins led to the induction of significantly increased antibody titres targeting the conserved CD4 binding site of Env despite similar overall antibody titres. These refocused antibody responses resulted in increased serum neutralising titres compared to animals receiving unmodified protein. Secondly, an epitope masking strategy was developed to reduce or eliminate the immunogenicity of neutralisation-irrelevant surfaces. This was achieved using site-selective addition of theoretically immunosilent glycoconjugates to lysine residues. Masking of model protein hen egg lysozyme (HEL) led to site-selective loss of antibody binding to the modification sites in vitro, which translated into refocusing of antibody responses from masked to unmasked epitopes in vivo. Mutant HIV-1 and influenza virus surface glycoproteins were designed that had lysine residues removed from close proximity to the respective broadly neutralising epitopes, but added throughout the remaining surface. Masking of these mutant proteins with second-generation glycoconjugates led to predictable perturbations of antibody binding in vitro. However, administration of these modified glycoproteins revealed unexpectedly that the masking glycans were highly immunogenic in vivo. Thus, this strategy may well prove useful if truly non-immunogenic glycoconjugates can be identified. Taken together, these chemical modifications of vaccine antigens may allow focused targeting of specific antigenic regions for increased B cell recognition, and may thus be a valuable tool for vaccine antigen design.

Failure to process prelamin A, by the enzyme ZMPSTE24, leads to the build up of farnesylated prelamin A, which has been implicated in causing the symptoms experienced in laminopathies and HIV therapy. A common feature to these conditions is the development of an irregular nuclear boundary, often including deep invaginations that form a nucleoplasmic reticulum. Additionally, dysregulated lipid synthesis is frequently associated with improper lamin A processing and I set out to address the molecular mechanisms behind these two features that could explain lipoatrophy experienced in patients. By using siRNA targeted against Zmpste24 I utilised an array of biochemical, molecular and imaging techniques to uncover a mechanism that leads to the production of a nucleoplasmic reticulum that was dependent on both the farnesylated tail of prelamin A and the phosphatidylcholine synthesising enzyme CCTα. The morphology of this structure consisted of an invagination of both the inner and outer nuclear membranes with a cytoplasmic core or just invagination of the inner nuclear membrane. Serial section dual axis electron tomography provided a new insight into the ultrastructural changes at the nuclear periphery that revealed novel structural features. The dysregulation of lipid synthesis was assessed by investigating the effects farnesylated prelamin A has on the distribution and dynamics of the transcription factor SREBP-1 and assessment of the downstream consequences this has on its targets that regulate adipocyte differentiation potential. Finally, the metabolomic profile of an HIV protease inhibitor that leads to prelamin A build up was generated and revealed increases in lipolysis, glycolysis and mediators of inflammation. The research presented offers a new insight into the development of a convoluted nuclear boundary and nucleoplasmic reticulum, in the context of lamin A mutants and how dysregulated lipid synthesis, caused by farnesylated prelamin A, leads to lipoatrophy.

Background: The 22q11 deletion syndrome (22q11DS), also known as DiGeorge Syndrome, affects ~1/5000 live born children. Congenital heart defects (typically outflow tract and interrupted aortic arch) are present in 75% of individuals with 22q11DS and are the major cause of mortality. Other defects are cleft palate, thymus hypoplasia, inner ear defects and neuropsychiatric abnormalities. Df(16)1 mice carry a ~1 Mb hemizygous deletion on mouse chromosome 16 in a region syntenic with 22q11 and phenocopies 22q11DS. TBX1 is a DNA-binding transcription factor located in this interval and is required for neural crest cell proliferation and migration and for cardiac development. TBX1 point mutations have been identified in patients with DiGeorge syndrome. Thus TBX1 is thought to be a major gene responsible for the cardiac phenotype in 22q11DS. A key unresolved issue is the mechanism of reduced penetrance of cardiac malformations. One possibility is environmental variation during cardiogenesis. A second possibility is that variation in the TBX1 protein interaction network results in variable penetrance of the phenotype. Mutations in TBX1 or interacting partners could affect the structure of this protein interaction network. Aim: The aim of this thesis is to characterize the molecular mechanism of TBX1 function using biochemical and genetic approaches and to define the role of environmental variation on the DiGeorge phenotype. Results First part. Interaction of Df(16)1 with high-fat maternal diet. To determine if a maternal high-fat diet affects the penetrance of cardiac and thymus malformations in the Df1 deletion mouse model, wild-type and Df1 heterozygous embryos from control and high-fat diet groups were analyzed. No significant difference in the penetrance or the severity of cardiac malformations between these groups was found. These results do not support the idea that change in the fat content of maternal diet affects phenotype in this model. Thus, it is possible that high-fat diet interacts specifically with left-right patterning rather than with the genetic control of pharyngeal arch development and neural crest cell migration and survival. Second part. George, a novel ENU induced mutation in Tbx1. The George mutation, identified and mapped to Chr16 between rs4161352 and D16Mit112, results in fully penetrant cleft palate, cardiac malformations (VSD, IAA, CAT), absent cochlea and abnormal semicircular canals, and absent thymus resembling the human DiGeorge phenotype. Tbx1 lies in this interval and sequencing identified a G > A point mutation in exon 3 which is predicted to cause a Arginine to Glutamine change at amino acid position 160. George fails to genetically complement a Tbx1 null allele, confirming that it is causative and that George is functionally a null allele. RT-PCR showed that the George mutation affects splicing, resulting in a transcript lacking exon 3. This causes the loss of 34 amino acids within the TBX1 T-box domain, thus predicting that it affects DNA binding. Transactivation assays show that while the R160Q amino acid substitution significantly reduces the transactivation capacity of TBX1, surprisingly the loss of exon 3 does not affect this function. Analysis of endogenous TBX1 in developing embryos by Western blot showed that the protein expression is absent or significantly reduced. This finding suggests that the observed George phenotype is caused primarily by a loss of TBX1 protein expression. Third part. Investigation of the protein interaction network surrounding TBX1. In order to get a better insight into the protein network surrounding TBX1, a TBX1 split renilla-luciferase protein complementation assay was set up which allowed to test the physical interaction between TBX1 and several putative interactors. It was found that GATA4, SMARCAD1, RBBP5 and PTDSR interact with wild-type TBX1 in HEK293T cells. The R160Q point mutation and the loss of exon 3 affect some of these interactions supporting the idea that variation in the protein interaction network may, at least in part, be responsible for the DGS phenotype.

Dissertation / The aim of this study was to determine the knowledge of persons with hypertension in a selected geographical area regarding cardiovascular risk factors in order to make recommendations for patient education. A quantitative, non-experimental, descriptive study was done in the form of a survey using a questionnaire as measuring instrument. The population was hypertensive patients from selected private medical practices in the western part of KwaZulu-Natal and the bordering eastern part of the Free State. Convenience sampling was used and 46 respondents participated in the study. Only 16 (35%) of the respondents achieved a percentage on or above the competency indicator of 50%. Respondents performed worst in questions where definitions, for example hypertension, were assessed. Recommendations for a patient education document, nursing practice and further research were made. / Health Studies / M.A. (Health Studies)

Neurodegeneration is a progressive loss of neuron function or structure, including death of neurons, and occurs at many different levels of neuronal circuitry. In this thesis I discuss Parkinson’s Disease (PD), the second most common neurodegenerative disease (NDD). PD is a devastating progressive NDD often with delayed diagnosis due to detection methods that depend on the appearance of visible motor symptoms. By the time cardinal symptoms manifest, 60 to 80 percent or more of the dopamine-producing cells in the substantia nigra are irreversibly lost. Although there is currently no cure, earlier detection would be highly beneficial to manage treatment and track disease progression. However, today’s clinical diagnosis methods are limited to subjective evaluations and observation. Onset, symptoms and progression significantly vary from patient to patient across stages and subtypes that exceed the scope of a standardized diagnosis. The goal of this thesis is to provide the basis of a more general approach to study the brain, investigating early detection method for NDD with focus on PD. It details the preliminary development, testing and validation of tools and methods to objectively quantify and extrapolate motor and non-motor features of PD from behavioral and cognitive output during everyday life. Measures of interest are categorized within three domains: the motor system, cognitive function, and brain activity. This thesis describes the initial development of non-intrusive tools and methods to obtain high-resolution movement and speech data from everyday life and feasibility analysis of facial feature extraction and EEG for future integration. I tested and validated a body sensor system and wavelet analysis to measure complex movements and object interaction in everyday living situations. The sensor system was also tested for differentiating between healthy and impaired movements. Engineering and design criteria of the sensor system were tested for usability during everyday life. Cognitive processing was quantified during everyday living tasks with varying loaded conditions to test methods for measuring cognitive function. Everyday speech was analyzed for motor and non-motor correlations related to the severity of the disease. A neural oscillation detection (NOD) algorithm was tested in pain patients and facial expression was analyzed to measure both motor and non-motor aspects of PD. Results showed that the wearable sensor system can measure complex movements during everyday living tasks and demonstrates sensitivity to detect physiological differences between patients and controls. Preliminary engineering design supports clothing integration and development of a smartphone sensor platform for everyday use. Early results from loaded conditions suggest that attentional processing is most affected by cognitive demands and could be developed as a method to detect cognitive decline. Analysis of speech symptoms demonstrates a need to collect higher resolution spontaneous speech from everyday living to measure speech motor and non-motor speech features such as language content. Facial expression classifiers and the NOD algorithm indicated feasibility for future integration with additional validation in PD patients. Thus this thesis describes the initial development of tools and methods towards a more general approach to detecting PD. Measuring speech and movement during everyday life could provide a link between motor and cognitive domains to characterize the earliest detectable features of PD. The approach represents a departure from the current state of detection methods that use single data entities (e.g.one-off imaging procedures), which cannot be easily integrated with other data streams, are time consuming and economically costly. The long-term vision is to develop a non-invasive system to measure and integrate behavioral and cognitive features enabling early detection and progression tracking of degenerative disease.

The purpose of this study was to develop a support programme for facilitating the integration of nutrition and food security with HIV prevention, treatment and care. The study was organised in three phases. Phase one was a quantitative cross-sectional survey that employed a structured interview with people living with HIV among selected two public hospitals and three health centres. The second phase employed focus group discussion with senior health experts to explore their perspective and experience in integrating nutrition and food security with HIV prevention, treatment, and care. The findings indicated that malnutrition and food insecurity were highly prevalent and significantly affected the treatment outcome and quality of life of PLWHA in the region. Socio-economic, clinical features and structural factors, such as educational status, place of residence, household income, source of drinking water, kind of toilet facility, inadequate dietary diversity, poor asset possession, opportunistic infections, duration on ART, CD4 cell count, and health system-related factors such as lack of viral and CD4 analysis laboratories and inconsistent antiretroviral medication supply were found as predictors of malnutrition and food insecurity. To cope up with the dire impact of malnutrition and food insecurity, short term, erosive and unsustainable food consumption coping strategies were employed. Based on the findings, the researcher developed a support programme for facilitating the integration of nutrition and food security with HIV prevention, treatment and care as phase three of the study. The developed programme is holistic and focuses on multi- and intersectoral collaboration to improve the treatment outcome, quality of life and overall wellbeing people living with HIV. / Health Studies / D. Litt. et Phil. (Health Studies)

Heart disease including ischemic heart disease is the highest contributor to death and morbidity in the western world. The studies presented were conducted to determine possible pathways of protection of the heart against ischemia/reperfusion. We employed adenovirus mediated over-expression of Methionine sulfoxide reductase A (MsrA) in primary neonatal rat cardiac myocytes to determine the effect of this enzyme in protecting against hypoxia/reoxygenation. Cells transfected with MsrA encoding adenovirus and subjected to hypoxia/reoxygenation exhibited a 45% decrease in apoptosis as compared to controls. Likewise total cell death as determined by levels of Lactate Dehydrogenase (LDH) release was dramatically decreased by MsrA overexpression. The initial hypothesis that led to our testing sulindac was based on the fact that the S epimer of sulindac was a substrate for MsrA and that this compound might function as a catalytic anti-oxidant based on a reaction cycle that involved reductio n to sulindac sulfide followed by oxidation back to sulindac. To test this we examined the protective effect of sulindac in hypoxia re-oxygenation in both cardiac myocytes in culture and using a Langendorff model of myocardial ischemia. Using this model of myocardial ischemia we showed that pre-incubation of hearts with sulindac, or the S and R epimers of sulindac resulted in protection against cell death. We present several lines of evidence that the protective effect of sulindac is not dependent on the Msr enzyme system nor does it involve the well established role of sulindac as a Cyclooxygenase (COX) inhibitor. Numerous signaling pathways have been implicated in myocardial protective mechanisms, many of which require fluctuations in ROS levels as initiators or mediators. / Sulindac shows very good potential as a preconditioning agent that could induce tissue protection against oxidative damage.Blocking of preconditioning pathways by administration of the PKC blocker chelerythine abrogated the ischemic protection afforded by sulindac. Secondly, an end-effector of preconditioning, inducible nitric oxide synthase (iNOS),was found to be induced by greater than 5 fold after 48 h prior feeding sulindac. / by Ian Moench. / Thesis (Ph.D.)--Florida Atlantic University, 2008. / Includes bibliography. / Electronic reproduction. Boca Raton, Fla., 2008. Mode of access: World Wide Web.

O lítio é freqüentemente utilizado no tratamento do Transtorno Bipolar, doença associada a um risco aumentado para demência. Evidências experimentais sugerem efeitos neuroproterores do lítio. O lítio inibe a amiloidogênese e a fosforilação da proteína tau tanto in vitro como in vivo. Estes são processos importantes na patogênese da doença de Alzheimer. O objetivo este estudo foi a investigação do efeito do lítio na prevalência de Transtorno Cognitivo Leve e doença de Alzheimer em 114 bipolares idosos eutímicos. Todos os sujeitos completaram uma avaliação catamnéstica, psicopatológica e cognitiva que incluía o mini-exame do estado mental (Mini-mental), o teste cognitivo de Cambridge (CAMCOG) e o questionário do informante sobre o declínio cognitivo do idoso (IQCODE). Foi feita uma comparação da prevalência de Transtorno Cognitivo Leve e doença de Alzheimer entre pacientes em uso de lítio e pacientes em uso de outros estabilizadores de humor. Os sujeitos que entraram na pesquisa tinham em média 68,2 ± 5,0 anos e preenchiam os critérios da Décima Revisão da Classificação Internacional de Doenças e Problemas de Saúde Relacionados (CID-10) para o transtorno bipolar. Durante a avaliação os bipolares estavam eutímicos. Eutimia foi definida como uma pontuação máxima de 7 pontos na escala de Hamilton de 21 pontos para Depressão e 4 na escala de Young para mania. 66 pacientes em uso contínuo do lítio por 6 anos em média foram comparados com 48 pacientes em tratamento com outros estabilizadores de humor. O diagnóstico de Transtorno Cognitivo Leve foi feito de acordo com os critérios de Petersen(1999) e de doença de Alzheimer de acordo com o critério do “National Institute for Communicative Disorders and Stroke - Alzheimer’s Disease and Related Disorders Association" (NINCDS/ADRDA). A prevalência de demência nesta amostra (19,4%) foi mais elevada do que o esperado para uma população comparável (7,1%). A prevalência de doença de Alzheimer entre aqueles com lítio foi 4,5% quando comparada com 33,3% entre aqueles sem lítio. Controlando idade e outras variáveis relacionadas ao curso da doença, o efeito do lítio na prevalência de doença de Alzheimer permaneceu significativo (OR = 0,079; p < 0,001). Nenhuma associação foi encontrada com Transtorno Cognitivo Leve. A alta da prevalência de doença de Alzheimer neste estudo está de acordo com as evidências de risco aumentado para demência em pacientes bipolares. Nesta amostra o tratamento com lítio reduziu a prevalência de Alzheimer aos níveis da população idosa em geral. Estes achados estão de acordo com os efeitos neuroprotetores do lítio em eventos cruciais para a patologia da doença de Alzheimer. Estudos prospectivos são necessários para avaliar se o lítio também pode ser efetivo na prevenção de doença de Alzheimer em outras populações. / Lithium is widely used in the treatment of bipolar disorder, a condition associated with an increased risk for dementia. Experimental evidence suggests that lithium has a neuroprotective effect. Both in vitro and in vivo, lithium inhibits amyloidogenesis and phosphorilation of tau protein, which are two crucial processes in the pathogenesis of Alzheimer’s disease. The objective of this study was to investigate the effect of lithium on the prevalence of Mild Cognitive Impairment and Alzheimer’s disease in 114 elderly euthymic bipolar patients. Subjects completed a thorough catamnestic, psychopathological and cognitive tests evaluation including the Mini-mental state evaluation, Cambridge cognitive test (CAMCOG) and the informant questionnaire on cognitive decline in the elderly (IQCODE). The prevalence of Mild Cognitive Impairment and Alzheimer’s disease between patients on lithium therapy and patients on treatment with other mood-stabilizing drugs was compared. Patients were 68.2 ± 5.0 years old and fulfilled of the International Classification of Diseases - 10th Revision (ICD-10) diagnosis for bipolar disorder. At the time of the evaluation patients were euthymic, as defined by a maximum score of 7 in the 21-item Hamilton Rating Scale for Depression, and 4 in the Young Mania Rating Scale. Sixty-six patients were continuously being treated with lithium for six years, on average, and 48 patients were receiving other mood-stabilizing drugs. Diagnosis of Mild Cognitive Impairment was made according to Petersen (1999) and of Alzheimer’s disease was made according to the National Institute for Communicative Disorders and Stroke - Alzheimer’s Disease and Related Disorders Association (NINCDS/ADRDA) criteria. The overall prevalence of dementia in our sample (19.4%) was higher than the prevalence expected in the age-comparable general population (7.1%). The prevalence of Alzheimer’s disease among lithium users was 4.5% as compared to 33.3% among non-users. After controlling for age and other variables related to the clinical course of the bipolar disorder, the effect of lithium on Alzheimer’s disease prevalence remained significant (OR = 0.079; p < 0.001). No association was found with Mild Cognitive Impairment. The higher prevalence of Alzheimer’s disease in our study supports the reports of increased risk for dementia in bipolar patients. In our sample, lithium treatment reduced the prevalence of Alzheimer’s disease to the levels of the general elderly population. This finding is in line with the neuroprotective effects of lithium on crucial events for the pathology of Alzheimer’s disease. Further prospective studies are needed to clarify whether lithium may also be effective in the prevention of Alzheimer’s disease in the general population.

A Doença de Alzheimer é uma doença neurodegenerativa progressiva e de início tardio, que compromete principalmente as areas da cognição, julgamento e estabilidade emocional. Esta doença se caracteriza por dois tipos de lesões cerebrais características: emaranhados neurofibrilares e placas senis. Os emaranhados neurofibrilares são compostos por uma proteína do citoesqueleto (proteína tau) hiperfosforilada e agregada. As placas senis são formadas por agregados da proteína &#946;-amilóide. A doença de Alzheimer é resultado dainteração de vários fatores ainda incompletamente elucidados; não obstante, o estresse oxidativo e os processos inflamatórios ocupam posição de destaque dentre esses fatores. Neste trabalho, avaliamos as atividades das enzimas eritrocitárias superóxido dismutase, catalase e glutationa peroxidase, assim como o conteúdo plasmático de glutationa total, vitamina C, &#945;-tocoferol, &#946;-caroteno, licopeno e coenzima Q10. A esses parâmetros antioxidantes foram contrapostas medidas de oxidação de lipídios e proteínas plasmáticas. Adicionalmente, efetuamos a avaliação das expressões monocitárias de HLADR e CD-11b, e das citocinas IL-6, IL-1&#945; e TNF-&#945;. Nossos resultados mostram que os pacientes de doença de Alzheimer possuem níveis circulantes de atocoferol inferiores aos pacientes controles, e possuem monócitos que apresentam maior expressão basal de HLA-DR e maior produção de IL-1&#945; quando estimulados por LPS. Esses resultados fortalecem a hipótese inflamatória na doença de Alzheimer, de acordo com trabalhos recentes que apontam bons resultados com o a-tocoferol na sua prevenção e tratamento. / Alzheimer\'s disease is a late-onset, progressive degenerative disease that affects mainly the judgement, emotional stability and memory domains. This disease is characterized by two telltale cerebral lesions: neurofibrillary tangles and senile plaques. Neurofibrillary tangles are constituted by hyperphosphorylated cytoskeleton protein tau aggregates, while senile plaques are mainly composed by &#946;-amyloid protein aggregates. Alzheimer\'s disease is the outcome of a complex interaction among several factors which are not fully understood yet; nevertheless it is clear thar oxidative stress and inflammatory pathways rate high among these factors. In this work, we evaluated the erythrocytic acitivities of superoxide dismutase, catalase and glutathione peroxidase, as well as the plasma levels of total glutathione, &#945;-tocopherol, &#945;-carotene, lycopene, and coenzyme Q10. These antioxidant parameters were confronted with plasmatic levels of protein and lipid oxidation products. Additionally, we measured the basal expression of monocyte HLA-DR and CD-11b, as well as the monocyte production of the cytokines IL1-&#945;, IL-6 and TNF-&#945;. Our results show that Alzheimer\'s Disease patients show lower plasmatic levels of &#945;-tocopherol when compared to control patients, and have higher basal monocyte HLA-DR expression associated with higher IL-1&#945; production when stimulated by LPS. These results lend support to the inflammatory theory of Alzheimer\'s disease, according to recent works that indicate good results of &#945;-tocopherol administration in both its prevention and treatment.