Doctor of Philosophy / The aim of the work presented in this thesis was to examine the associations between the kinematics of the knee characterised by the tibiofemoral contact pattern, and degenerative change, in the context of anterior cruciate ligament (ACL) injury. While the natural history of degenerative change following knee injury is well understood, the role of kinematics in these changes is unclear. Kinematics of the knee has been described in a variety of ways, most commonly by describing motion according to the six degrees of freedom of the knee. The advantage of mapping the tibiofemoral contact pattern is that it describes events at the articular surface, important to degenerative change. It was hypothesised that the tibiofemoral contact pattern would be affected by injury to the knee. A model of ACL injury was chosen because the kinematics of the knee have been shown to be affected by ACL injury, and because the majority of chronic ACL-deficient knees develop osteoarthritis, the associations between kinematics and degenerative change could be explored. A technique of tibiofemoral contact pattern mapping was established using MRI, as a quantifiable measure of knee kinematics. The tibiofemoral contact pattern was recorded from 0º to 90º knee flexion while subjects performed a leg-press against a 150N load, using sagittal magnetic resonance imaging (MRI) scans. The technique was tested and found to be reliable, allowing a description of the tibiofemoral contact pattern in 12 healthy subjects. The tibiofemoral contact patterns of knee pathology were then examined in a series of studies of subjects at a variety of stages of chronicity of ligament injury and osteoarthritis. Twenty subjects with recent ACL injury, 23 subjects with chronic ACL deficiency of at least 10 years standing, and 14 subjects with established osteoarthritis of the knee were recruited. The 20 subjects with recent ACL injury were examined again at 12 weeks and 2 years following knee reconstruction. The tibiofemoral contact patterns were examined for each group of subjects and the associations between changes in the contact patterns and evidence of joint damage explored. Evidence of joint damage and severity of osteoarthritis were recorded from xrays, diagnostic MRI, operation reports and bone densitometry at the tibial and femoral condyles of the knee. Each of the three groups with knee pathology exhibited different characteristics in the tibiofemoral contact pattern, and these differences were associated with severity of joint damage and osteoarthritis. The recently ACL-injured knees demonstrated a tibiofemoral contact pattern that was posterior on the tibial plateau, particularly in the lateral compartment. Those with chronic ACL deficiency demonstrated differences in the contact pattern in the medial compartment, associated with severity of damage to the knee joint. Osteoarthritic knees showed reduced femoral roll back and longitudinal rotation that normally occur during knee flexion. Two years following knee reconstruction there was no difference between the contact pattern of the reconstructed and healthy contralateral knees. This technique of tibiofemoral contact pattern mapping is sensitive to the abnormal characteristics of kinematics in ligament injury and osteoarthritis. This is the first time the tibiofemoral contact characteristics of chronic ACL-deficient and osteoarthritis knees have been described and links examined between tibiofemoral contact patterns and degenerative change.

Doctor of Philosophy / Nineteenth century philosophy and anatomy regarded the nervous system as the only pathway of communication between the brain and body but now, research in the field of psychoneuroimmunology (PNI) has provided evidence to prove the age-old belief that there is a connection between the mind (or mental/emotional states) and the body. Researchers in PNI have now shown that the communication between the nervous and immune systems is bi-directional – i.e. there is a psychological reaction to physical disease and a somatic presentation of psychological disorders - and that the immune system, the autonomic nervous system, the endocrine system and the neuropeptide systems all communicate with each other by means of chemicals called messenger molecules or ligands. This paper outlines research into the treatment of Irritable Bowel Syndrome (IBS) with hypnotherapy, taking into account the mind-body connection and treating both the patient’s physiological and emotional/psychological symptoms rather than treating the physiological symptoms only. In other words, using a more holistic approach to the treatment of IBS. IBS is probably the most common functional gastrointestinal disorder encountered by both gastroenterologists and physicians in primary care. It is estimated that from 10% to 25% of the general population suffer from this condition and that it comprises about 30-50% of the gastroenterologists’ workload, yet the aetiology of IBS is unknown and, so far, there is no cure. Researchers are beginning to view IBS as a multi-faceted disorder in which there appears to be a disturbance in the interaction between the intestines, brain, and autonomic nervous system, resulting in an alteration in the regulation of bowel motility and/or sensory function. Most researchers agree that a subset of IBS sufferers have a visceral hypersensitivity of the gut or, more specifically, an increased perception of sensations in the gut. To date, studies of IBS have proposed previous gastroenteritis, small intestine bacterial overgrowth, psychosocial factors, a genetic contribution, and an imbalance of neurotransmitters as either possible causes or playing a part in the development of IBS. It is generally agreed that a patient’s emotional response to stress can exacerbate the condition. In section 1 of the thesis, the introduction, a detailed description and background appropriate to the study undertaken are provided, including aspects of epidemiology, diagnostic symptom criteria and clinical relevance of the Irritable Bowel Syndrome. Previous studies of various forms of treatment for IBS are discussed with the main emphasis being on treatment with hypnotherapy. All these therapies have concentrated on either mind or body treatments whereas this study demonstrates how hypnotherapy, and the use of imagery, addresses both mind and body. Finally, the rationale for the current study and the specific aims of the thesis are outlined. In section 2, the methodology and assessment instruments used in the clinical trial are discussed, as well as recruitment processes, research plan and timetable, and treatment schedule. Statistical analyses are provided and the main outcomes measures of the clinical trial, its limitations and scientific implications are addressed.

Doctor of Philosophy / The aim of the work presented in this thesis was to examine the associations between the kinematics of the knee characterised by the tibiofemoral contact pattern, and degenerative change, in the context of anterior cruciate ligament (ACL) injury. While the natural history of degenerative change following knee injury is well understood, the role of kinematics in these changes is unclear. Kinematics of the knee has been described in a variety of ways, most commonly by describing motion according to the six degrees of freedom of the knee. The advantage of mapping the tibiofemoral contact pattern is that it describes events at the articular surface, important to degenerative change. It was hypothesised that the tibiofemoral contact pattern would be affected by injury to the knee. A model of ACL injury was chosen because the kinematics of the knee have been shown to be affected by ACL injury, and because the majority of chronic ACL-deficient knees develop osteoarthritis, the associations between kinematics and degenerative change could be explored. A technique of tibiofemoral contact pattern mapping was established using MRI, as a quantifiable measure of knee kinematics. The tibiofemoral contact pattern was recorded from 0º to 90º knee flexion while subjects performed a leg-press against a 150N load, using sagittal magnetic resonance imaging (MRI) scans. The technique was tested and found to be reliable, allowing a description of the tibiofemoral contact pattern in 12 healthy subjects. The tibiofemoral contact patterns of knee pathology were then examined in a series of studies of subjects at a variety of stages of chronicity of ligament injury and osteoarthritis. Twenty subjects with recent ACL injury, 23 subjects with chronic ACL deficiency of at least 10 years standing, and 14 subjects with established osteoarthritis of the knee were recruited. The 20 subjects with recent ACL injury were examined again at 12 weeks and 2 years following knee reconstruction. The tibiofemoral contact patterns were examined for each group of subjects and the associations between changes in the contact patterns and evidence of joint damage explored. Evidence of joint damage and severity of osteoarthritis were recorded from xrays, diagnostic MRI, operation reports and bone densitometry at the tibial and femoral condyles of the knee. Each of the three groups with knee pathology exhibited different characteristics in the tibiofemoral contact pattern, and these differences were associated with severity of joint damage and osteoarthritis. The recently ACL-injured knees demonstrated a tibiofemoral contact pattern that was posterior on the tibial plateau, particularly in the lateral compartment. Those with chronic ACL deficiency demonstrated differences in the contact pattern in the medial compartment, associated with severity of damage to the knee joint. Osteoarthritic knees showed reduced femoral roll back and longitudinal rotation that normally occur during knee flexion. Two years following knee reconstruction there was no difference between the contact pattern of the reconstructed and healthy contralateral knees. This technique of tibiofemoral contact pattern mapping is sensitive to the abnormal characteristics of kinematics in ligament injury and osteoarthritis. This is the first time the tibiofemoral contact characteristics of chronic ACL-deficient and osteoarthritis knees have been described and links examined between tibiofemoral contact patterns and degenerative change.

La dysarthrie dans la maladie de Parkinson (MP) fait partie des symptômes qui ne répondent pas de manière satisfaisante aux prises en charge médicales. Bien que largement étudiée, certains aspects de la dysarthrie restent insuffisamment détaillés : quels sont ses corrélats physiologiques, ses liens avec des fonctions non-motrices et ses répercussions fonctionnelles ? Construit en quatre axes, ce projet doctoral avait pour objectif d'apporter des éléments de réponse à ces questionnements.Un premier axe a exploré les bases physiopathologiques de la dysarthrie. Considérant les effets du traitement médicamenteux et de la neurochirurgie, nous avons montré qu'aucun de ces traitements ne permettait de restaurer un profil neuro-fonctionnel similaire à celui de participants sains, d'autant plus lors d'une tâche combinant motricités axiale et distale. Le second axe a porté sur les liens entre dysarthrie et fonctions non-motrices. Nos résultats suggèrent que ces deux catégories de fonctions pourraient évoluer conjointement, mais aussi se majorer mutuellement. À l'aide de comparaisons inter-dysarthrie et inter-linguistique, le troisième axe a considéré l'impact fonctionnel de la dysarthrie. Nous avons rapporté que les conséquences de la dysarthrie ne sont pas fonction de l'intelligibilité, qu'elle affecte davantage les patients ayant un profil akinéto-rigide et qu'elle réduit les différences inter-linguistiques. L'étude des rééducations orthophoniques a constitué notre axe quatre. Nos conclusions ont relevé que pour être efficaces, les rééducations doivent être focalisées sur un unique paramètre de parole, être réalisées de manière intensive et apporter un retour au patient. / Dysarthria in Parkinson’s disease (PD) is one of the symptoms that do not respond well to treatments. Even though it has been widely described, some aspects of dysarthria remain insufficiently detailed: what are its physiological correlates, its relations with non-motor functions and its functional consequences? Structured in four axes, this thesis aimed at bringing some elements to address these issues.The first axis explored the pathophysiology of dysarthria. Considering the effects of pharmacological and neurosurgical treatments, we demonstrated that none of these treatments allowed to restore a neuro-functional profile equivalent to healthy controls, especially during a task combining axial and distal actions. The second axis focused on the relations between dysarthria and non-motor functions. Some outcomes suggested that these two categories of functions could progress jointly, but also reinforce each other. Using between-dysarthria and between-linguistic comparisons, the third axis considered the functional consequences of dysarthria. We reported that the consequences of dysarthria do not depend on the intelligibility degradation, that it impacts more the patients with an akineto-rigid profile, and that it tends to weaken the between-linguistic differences. The study of speech therapies represented our fourth axis. We concluded that, to be effective, speech therapies in PD have to focus on a unique speech parameter, to be realised intensively and to give feedback to the patient on their productions.

Oxygen is one of the most important molecules in human beings. Our research is focused on how the human body can respond and adapt to the physiological challenge posed by a lack of oxygen. Ascorbic acid (Vitamin C) is one of the most important and considered the most effective water-soluble, chain-breaking antioxidant in human plasma, with the capacity to prevents damage by free radicals. This thesis presents four studies investigating the phenomenon of Reactive Oxygen Species (ROS) generation in the many different surgical conditions in the animal and in the human. Study one investigated the geometry and thermodynamic properties of vitamin C. Calculations were carried out at the restricted and unrestricted B3LYP/6-31+G(d,p), B3LYP/6-311++G(d,p) and B3LYP/EPR-II levels for two conformers (1 and 2) of L-ascorbic acid and their respective oxidation products to monodehydroascorbates of ab-initio methods by Gaussian O3W package. Conformer 1, free radical properties are compared with previously published calculations in the gaseous and aqueous solution states and with experimental EPR values. Calculated molecular structures, EPR (electron paramagnetic resonance spectroscopy), the vibration spectral and energetic properties and all are reported including some proposed changes to previous EPR assignments. Conformer 2 of L-ascorbic acid is predicted to have lower energy than Conformer 1, under the method and basis sets used, by between 11 and 26 kJ mol-1 and is stabilised by internal hydrogen bonding. Relaxed potential energy surface (PES) scans were carried out for two proton transfer processes and relative energies of stable minima and barriers between them determined. Hydrogen transfer is predicted in two systems with favourable spatial arrangements of O–H and O groups for which relaxed potential energy surface scans are reported. Calculated vibrational wavenumber values are provided for selected C=C, C=O, C–H and O–H modes assigned to particular groups and significant calculated EPR hyperfine coupling constants (HCC) values for splitting by H(1) and C(13) for radical species are also reported. These calculations contribute to a better understanding of the complex role of L-ascorbic acid and its various oxidised, neutral, ionic and radical forms in biochemistry and medicine. Study two examined if vitamin C could ameliorate the damaging effects of I-R on myocardium and we postulated that the mechanism of vitamin C protection against iii I-R-induced cell death involved quenching of ROS. In the vitamin C group after 5 min of reperfusion a significant, sudden increase of diastolic pressure in the heart was noted and reached a maximum of 77 mmHg after 12 min of reperfusion and then gradually decreased to 51 mmHg after 60 min of reperfusion period but was quicker than in Control group reaching 37 mmHg by the end of the reperfusion period. The level of A·− (ascorbate free radicals) sudden and massive increased at the time of reperfusion in the Vitamin C group. This increase was associated with poor mechanical function in hearts as indicated by the significantly depressed recovery process. After 30 min of global, now-flow ischaemia and 60min of reperfusion infarct size averaged 33% ± 1 in Control group and 30 % ± 1 in Vitamin C group, respectively, (P<0.05). There is strong evidence that oxygen centered radicals contribute to postischaemic dysfunction after global ischaemia. Our data unquestionably suggest that the large production of A·− was associated with a greater depression in myocardial contractile function, therefore could represent a marker of oxidative stress during I-R and could be related to the functional impairment during reperfusion. In summary, we have used the animal models of isolated heart perfusion to provide evidence that vitamin C did not reduce the infarct size, however “tendency” towards a decrease (↓) in infarct size with ascorbate and it protects from oxidative damage during global I-R as manifested by decreased concentrations of A·− and enhance recovery of mechanical function such as diastolic pressure and LVDP in postischaemic working rat hearts. Study three was designed to test the hypothesis that the physiological trauma associated with venous cannulation may artefactually stimulate systemic free radical formation in the acute phase that if not accounted for may under-estimate the oxidative stress response to exercise. The relationship between the time of venepuncture and the level of free radical generation during normoxic conditions was further investigated. The venous cannulation in Phase I, increased plasma A·− by 347 ± 173 AU/√G, P <0.05 after 2min of venepuncture with further increases observed after 5min and 10min of venous cannulation, respectively (403 ± 178 AU/√G; 462 ± 93 AU/√G, P < 0.05) vs baseline point time. After this time the level of A·− slightly blunted as to achieve a similar level to baseline point control after 30 minutes. In phase II the exerciseinduced increase in A·− was subsequently shown to be 48% greater (30min as opposed to the 2min post-cannulation resting baseline)(1754 ± 361 vs. 1979 ± 375 AU, P <0.05). Our findings demonstrate and confirm that venous cannulation per se stimulates iv the systemic formation of free radicals as an acute phase response which peaks at 10min and require approximately 15min to normalise. This has important interpretive implications for future studies that employ catheterisation. The final Study examined if the combination of exercise and inspiratory hypoxia would further compound regional tissue de-oxygenation that is frequently encountered during the ischaemic phase of surgery and thus, by consequence increase oxidative stress. The aim of the study was to further understand a potential relationship between oxidative stress and alterations in muscle oxygenation. Clear significant increases in the plasma concentration of A·− were detected in the peripheral blood of patients (normoxia(baseline) vs 6 data points of reperfusion after 5min of global ischaemic condition, P<0.05),(baseline vs immediate after ischaemia; 2337±525 vs 2633±508, AU, respectively). During global ischaemia the regional muscle oxygenation significantly decreased (↓∆O2Hb-oxyhaemoglobin), ↑∆HHb- deoxyhaemoglobin ), although increased regional blood volume (↑∆tHb- total haemoglobin). From the end of global ischaemia to 10 min after the regional muscle oxygenation progressively back to the start data point (↓∆HHb, ↑∆O2Hb). This study demonstrates for the first time that the I-R has got a big influence on the muscle oxygenation to increased ROS and the return of values towards baseline period in reperfusion stage appears to coincide with increased oxidative stress. Moreover, the present study has also demonstrated increased A·− level as early as the ischaemic phase of experiment independent of perioperative changes in the partial pressure of oxygen (pO2), elucidate a potentially important role for oxidative stress in provoking an appropriate vasodilation (NO-bioavailability) during the I-R period. This work demonstrates that; - Ascorbate is an antioxidant that can scavenge tissue and blood borne free radical, is essential in controlled amounts and is capable of initiating protective adaptation in the face of oxidative stress for the maintenance of physiological homeostasis. - Reperfusion is always associated with a sudden and massive release of ascorbate free radicals, with a maximal liberation within the first minutes of reperfusion. Vitamin C tended to reduce infarct size and protects from oxidative damage during global ischaemia and reperfusion. - The venous cannulation alone is enough per se stimulates the systemic formation of free radicals as a acute phase response. If this baseline artefact is not taken into account, the true magnitude of the exercise-induced oxidative stress response will be under-estimated. / The I-R has got a major influence on the muscle oxygenation to increased ROS and the return of values towards baseline period in reperfusion stage appears to coincide with increased oxidative stress. Using the state-of-the-art molecular techniques that include Electron Paramagnetic Spectroscopy (EPR) for the direct detection of free radicals and Near Infrared Spectroscopy (NIRS) for the direct detection of muscle oxygenation these studies have attempted to translate the basic mechanisms associated with free radical formation during I-R and have provided unique insight into the basic mechanisms responsible for the oxidative stress with the ultimate objective of developing novel antioxidant interventions that can provide effective prophylaxis.

xii, 98 p. : ill. (some col.) / The short and long term pathophysiology of traumatic brain injury (TBI) has not been fully elucidated. Individuals recently suffering a mild TBI (mTBI) or having a history of TBI frequently suffer deficits in their ability to maintain and allocate attention within and between tasks. This dissertation examines the influence of mild and chronic TBI on performance of task switching. We employed spatial and numerical task switching paradigms to assess the behavioral deficits in mTBI, and we used an internally generated switching and an externally cued switching task along with functional Magnetic Resonance Imaging (fMRI) to assess the long term deficits in executive function resulting from chronic TBI. In the first experiment, individuals with mTBI were identified and tested within the first 48 hours of injury and then at a set interval 5, 14, and 28 days post injury. In the second investigation, individuals with chronic TBI were tested at least 12 months after their most recent injury. Healthy gender, age, and education matched controls were also tested in both studies. This research demonstrated that mTBI subjects display deficits in switching behavior within 48 hours of injury that failed to resolve a month post-injury; however, these costs did not generalize across the switching task types. Chronic TBI subjects performed internally generated and externally cued switching paradigms with a degree of success equivalent to that of healthy controls but displayed larger amounts of activation and recruited more areas of the brain at lower levels of difficulty and did not increase recruitment in a stepwise fashion at higher levels of difficulty. Mild TBI causes significant deficits in task switching, but there is specificity in these deficits. Chronic TBI patients performed at a level equivalent to that of controls but displayed different patterns and degree of activation. Taken together, these findings indicate that there may be a specific time frame during which task switching shows behavioral deficits, after which the subject may compensate for these deficits to produce normalized performance. / Committee in Charge: Dr. Paul van Donkelaar, Chair; Dr. Li-Shan Chou; Dr. Ulrich Mayr; Dr. Marjorie Woollacott

A anemia falciforme (AF) é a doença hereditária monogênica mais prevalente no Brasil, caracterizada pelo alto índice de morbimortalidade. Uma mutação de ponto no gene da globina beta da hemoglobina é a causa da doença. Características genéticas dos indivíduos além da possível heterogeneidade das moléculas associadas à hemólise e vasculopatia são responsáveis por uma variedade de manifestações e complicações clínicas. Os tratamentos disponíveis atualmente consistem no objetivo de amenizar as manifestações clínicas e reduzir o número de crises para uma melhor qualidade de vida destes pacientes. Considerando a importância dos MicroRNAs na regulação da expressão gênica e na fisiopatologia de diversas doenças, este estudo tem por objetivo a elucidação do mecanismo de ação destes potenciais reguladores na fisiopatologia da AF. Caracteriza-se por um estudo prospectivo comparativo, do tipo de pesquisa clínica transversal. Foram incluídos neste estudo 50 indivíduos, dos quais, 25 indivíduos normais sem a patologia, doadores do banco de sangue do Hospital de Clinicas de Porto Alegre (HCPA), e 25 pacientes homozigóticos SS, em acompanhamento médico no Centro de Referência em Doença Falciforme do HCPA A obtenção dos dados se deu pela reação da polimerase em cadeia em tempo real, com a seleção de quatro microRNAs candidatos selecionados de acordo com a predição de suas funções alvo já disponíveis na literatura (hsa-mir-15a, hsa-mir-210, hsa-mir-144 e hsa-mir-223). Foram comparadas as diferenças dos perfis de expressão de cada microRNA com a média do grupo controle, além das correlações entre as variáveis hematológicas, bioquímicas e manifestações clínicas, com a finalidade de avaliar a influência entre as variáveis positivamente ou negativamente. Resultados: Três dos quatro microRNAs tiveram seus níveis de expressão estatisticamente significativos em relação ao grupo controle (mir- 15a, mir-210 e mir-223). As correlações positivas identificadas foram do microRNA 15a com o microRNA 144, do microRNA 210 com o microRNA 223, além do microRNA 223 com as manifestações de úlceras. As correlações negativas identificadas foram do microRNA 15a em relação às plaquetas e síndrome torácica aguda, e do microRNA 144 em relação aos reticulócitos. Conclusão: Tal conhecimento poderá possibilitar estabelecer novos tratamentos e possíveis abordagens terapêuticas através do controle da expressão de genes específicos e sua interação direta com RNAs alvo. / Sickle cell anemia (FA) is the most prevalent monogenic hereditary disease in Brazil; it is characterized by variable and sometimes severe symptoms and high morbi-mortality. A point mutation of the beta globin gene is a cause of the disease. Genetic characteristics of individuals and the heterogeneous possibility of molecules associated with hemolysis and vasculopathy are responsible for the variability of clinical manifestations. The available treatments are aimed at mitigating the clinical manifestations and reducing the number of crisis for a better quality of life of these patients. This study aims to elucidate the mechanism of action of regulatory molecules in the pathophysiology of FA. It is characterized by a prospective comparative study, type of cross-sectional clinical research. Fifty individuals, 25 normal subjects, from the blood bank of the Hospital de Clínicas de Porto Alegre (HCPA), and 25 homozygous SS patients, from the Reference Center on Sickle Disease of HCPA. Real-time polymerase chain reaction measuring four microRNAs selected according to the predictions of their target functions in the literature (hsa-mir-15a, hsa-mir-210, Hsa-mir -144 and hsa-mir-223) Differences in the expression profiles of each microRNA with a mean of the control group were compared, as well as the correlations between hematological, biochemical and clinical manifestations, with the purpose of evaluating positive or negative influence between variables. Results: Three of the four microRNAs had their expression levels statistically significant in relation to the control group (mir- 15a, mir-210 and mir-223). A positive correlation was identified between microRNA 15a with the microRNA 144, the microRNA 210 with the microRNA 223, and microRNA 223 with positively correlated with leg ulcers. As for negative correlation we identified for microRNA 15a in relation to platelets and acute thoracic syndrome, and for microRNA 144 in relation to reticulocytes. Conclusion: Such knowledge may enable new treatments and possible therapeutic approaches by controlling the expression of specific genes and their direct interaction with target RNAs.

Patent foramen ovale (PFO) has been associated with an increased risk of decompression sickness (DCS) in divers. Pathophysiologicaly this has been ascribed to paradoxical embolization of nitrogen bubbles from venous blood to systemic circulation, resulting in obstruction of peripheral capillaries and ischemic injury. However, the role of PFO has been largely debated and experimental and prospective clinical data has been missing. It is of note, that this hypothesis is not only of theoretical importance. The proof of PFO as a causative factor of DCS and, importantly, of unpredictable events (unprovoked DCS) could affect millions of divers worldwide through improved therapy and prevention. In our research we aimed to describe the pathophysiological role of PFO in decompression sickness and to determine whether the prevention of arterialization of post-dive venous gas emboli (VGE) would decrease the incidence of unprovoked DCS in divers. We have screened 489 scuba divers for the presence of PFO by means of transcranial color-coded Doppler ultrasonography. In a retrospective analysis we found that the incidence of unprovoked decompression sickness was 7% among these divers and that PFO was the only risk factor. Subsequently, we have studied the occurrence of VGE and arterial gas emboli (AGE) in divers with...

A anemia falciforme (AF) é a doença hereditária monogênica mais prevalente no Brasil, caracterizada pelo alto índice de morbimortalidade. Uma mutação de ponto no gene da globina beta da hemoglobina é a causa da doença. Características genéticas dos indivíduos além da possível heterogeneidade das moléculas associadas à hemólise e vasculopatia são responsáveis por uma variedade de manifestações e complicações clínicas. Os tratamentos disponíveis atualmente consistem no objetivo de amenizar as manifestações clínicas e reduzir o número de crises para uma melhor qualidade de vida destes pacientes. Considerando a importância dos MicroRNAs na regulação da expressão gênica e na fisiopatologia de diversas doenças, este estudo tem por objetivo a elucidação do mecanismo de ação destes potenciais reguladores na fisiopatologia da AF. Caracteriza-se por um estudo prospectivo comparativo, do tipo de pesquisa clínica transversal. Foram incluídos neste estudo 50 indivíduos, dos quais, 25 indivíduos normais sem a patologia, doadores do banco de sangue do Hospital de Clinicas de Porto Alegre (HCPA), e 25 pacientes homozigóticos SS, em acompanhamento médico no Centro de Referência em Doença Falciforme do HCPA A obtenção dos dados se deu pela reação da polimerase em cadeia em tempo real, com a seleção de quatro microRNAs candidatos selecionados de acordo com a predição de suas funções alvo já disponíveis na literatura (hsa-mir-15a, hsa-mir-210, hsa-mir-144 e hsa-mir-223). Foram comparadas as diferenças dos perfis de expressão de cada microRNA com a média do grupo controle, além das correlações entre as variáveis hematológicas, bioquímicas e manifestações clínicas, com a finalidade de avaliar a influência entre as variáveis positivamente ou negativamente. Resultados: Três dos quatro microRNAs tiveram seus níveis de expressão estatisticamente significativos em relação ao grupo controle (mir- 15a, mir-210 e mir-223). As correlações positivas identificadas foram do microRNA 15a com o microRNA 144, do microRNA 210 com o microRNA 223, além do microRNA 223 com as manifestações de úlceras. As correlações negativas identificadas foram do microRNA 15a em relação às plaquetas e síndrome torácica aguda, e do microRNA 144 em relação aos reticulócitos. Conclusão: Tal conhecimento poderá possibilitar estabelecer novos tratamentos e possíveis abordagens terapêuticas através do controle da expressão de genes específicos e sua interação direta com RNAs alvo. / Sickle cell anemia (FA) is the most prevalent monogenic hereditary disease in Brazil; it is characterized by variable and sometimes severe symptoms and high morbi-mortality. A point mutation of the beta globin gene is a cause of the disease. Genetic characteristics of individuals and the heterogeneous possibility of molecules associated with hemolysis and vasculopathy are responsible for the variability of clinical manifestations. The available treatments are aimed at mitigating the clinical manifestations and reducing the number of crisis for a better quality of life of these patients. This study aims to elucidate the mechanism of action of regulatory molecules in the pathophysiology of FA. It is characterized by a prospective comparative study, type of cross-sectional clinical research. Fifty individuals, 25 normal subjects, from the blood bank of the Hospital de Clínicas de Porto Alegre (HCPA), and 25 homozygous SS patients, from the Reference Center on Sickle Disease of HCPA. Real-time polymerase chain reaction measuring four microRNAs selected according to the predictions of their target functions in the literature (hsa-mir-15a, hsa-mir-210, Hsa-mir -144 and hsa-mir-223) Differences in the expression profiles of each microRNA with a mean of the control group were compared, as well as the correlations between hematological, biochemical and clinical manifestations, with the purpose of evaluating positive or negative influence between variables. Results: Three of the four microRNAs had their expression levels statistically significant in relation to the control group (mir- 15a, mir-210 and mir-223). A positive correlation was identified between microRNA 15a with the microRNA 144, the microRNA 210 with the microRNA 223, and microRNA 223 with positively correlated with leg ulcers. As for negative correlation we identified for microRNA 15a in relation to platelets and acute thoracic syndrome, and for microRNA 144 in relation to reticulocytes. Conclusion: Such knowledge may enable new treatments and possible therapeutic approaches by controlling the expression of specific genes and their direct interaction with target RNAs.

(summary of doctoral dissertation) It has become increasingly apparent in recent years that there are important differences of many cardiovascular disorders including ventricular tachycardias in men and women. Gender differences have been observed in the epidemiology, pathogenesis and clinical presentation of various ventricular arrhythmias. Physiological menopause occurs as a part of a woman's normal aging process being based on the natural cessation of estradiol and progesterone production by the ovaries. The dramatic fall in circulating estrogens levels at menopause impacts many tissues including cardiovascular system. Because the incidence of coronary heart disease (CHD) rises significantly after menopause, it has been hypothesized that women's CHD advantage before menopause (in comparison to men of the same age) could be due to the protective effects of estrogens. However, controversial results have been reported since early nineties until today. While some studies found reduction in the incidence of CHD and in mortality from cardiovascular diseases some other studies failed to provide any evidence for an independent role of estradiol levels in determining CHD in postmenopausal women and some studies even found positive association of endogenous estradiol with the risk of CHD among women above...