O objetivo do presente estudo foi avaliar o papel do treinamento físico aeróbio, resistido ou combinado (aeróbio+resistido) no desenvolvimento do diabetes tipo 2 analisando mecanismos associados às complicações no diabetes em camundongos fêmeas com deficiência na produção leptina (ob/ob). Para tanto, foram utilizadas camundongos fêmeas, inicialmente com 4 semanas de idade, divididas em 6 grupos: ob/ob sedentárias com 4 semanas de vida (OS-4), selvagens sedentárias (SS) ou ob/ob sedentárias (OS-12) acompanhadas até a 12ª semana de vida, ob/ob treinamento aeróbio (OA), ob/ob treinamento resistido (OR) e ob/ob treinamento combinado (OC). Os grupos treinados foram submetidos a 8 semanas de treinamento físico dinâmico aeróbio em esteira (50 a 60% da velocidade máxima do teste de esforço) ou resistido em escada (4060% da carga máxima) ou a associação dos dois treinos (combinado). Foram avaliados: peso corporal; glicose, triglicérides e colesterol total sanguíneos; pressão arterial (PA) e frequência cardíaca (FC); sensibilidade barorreflexa (SBR); modulação autonômica cardiovascular; marcadores inflamatórios e hormonais; e parâmetros de estresse oxidativo. Os animais obesos (OS-12) apresentaram aumento de peso corporal, tecido adiposo, de glicemia, de triglicérides e de intolerância à glicose quando comparado aos animais selvagens (SS). Adicionalmente, o grupo OS-12 apresentou piores resultados nos testes aeróbio e de força. Não observamos diferenças entre os grupos SS e OS-12 em relação a PA e FC, porém o grupo OS-12 apresentou diminuição da variabilidade da frequência cardíaca (VFC) (33 ± 4ms2) e da sensibilidade barorreflexa em relação ao grupo SS (VFC: 178 ± 19 ms2). O grupo OS-12 apresentou aumento de angiotensina 2 nos tecidos renal e cardíaco, diminuição da adiponectina e aumento de citocinas inflamatórias no tecido adiposo e no baço em relação ao grupo SS. Somado a isso, os animais obesos apresentam maior dano a proteínas e lipoperoxidação e diminuição das enzimas antioxidantes em tecido renal e cardíaco em relação ao grupo SS. A comparação entre os grupos OS-4 e OS-12 evidenciou aumento de peso corporal, tecido adiposo, glicemia, intolerância à glicose e de parâmetros de estresse oxidativo no grupo OS-12 em relação ao grupo OS-4. A redução na VFC e na SBR foi observada no grupo OS-4 e no grupo OS-12. O treinamento físico por sua vez, diminuiu o ganho de peso e reduziu a glicemia e a intolerância à glicose nos três grupos treinados em comparação ao grupo OS-12. O treinamento físico aeróbio (61 ± 8ms2 e 6 ± 4 mmHg2) e resistido (66 ± 16ms2 e 6 ± 1,4mmHg2) foram eficientes em aumentar a VFC e diminuir a banda de baixa frequência da PA (simpático vascular) em relação ao grupo OS-12, porém o grupo OC (43 ± 7ms2 e 8 ± 0,9mmHg2) foi semelhante ao grupo OS-12 (10 ± 1,1mmHg2) e aos grupos OA e OR. Além disso, as três modalidades melhoraram a SBR. Os três tipos de treinamento reduziram os níveis de angiotensina 2 e aumentaram os níveis de angiotensina 1-7 em tecido adiposo, rim e coração. O treinamento físico aeróbio foi mais eficiente em melhorar o perfil inflamatório em tecido adiposo e no baço, uma vez que os grupos OA e OC apresentaram aumento de adiponectina e apenas o grupo OC apresentou diminuição de IL-6 e PAI-1 em relação ao grupo OS-12. Em relação ao estresse oxidativo, os três grupos treinados apresentaram diminuição de marcadores de lesão. Concluindo, nossos achados confirmam o desenvolvimento de disfunção metabólica ao longo da vida de camundongos ob/ob. É interessante notar que com 4 semanas de vida camundongos ob/ob apresentaram uma expressiva redução dos parâmetros da VFC. Este desbalanço autonômico, poderiam estar ocorrendo não só no coração, mas para outros tecidos, como o baço e o tecido adiposo, favorecendo a liberação de citocinas inflamatórias que poderiam induzir a longo prazo lesão de órgão alvo, como observado no presente estudo em coração e rins, por aumento de estresse oxidativo. Os grupos treinados, independente da modalidade, apresentaram melhora metabólica e na regulação autonômica cardiovascular, a qual foi acompanhada de alterações favoráveis no sistema renina-angiotensina, em mediadores inflamatórios e no perfil de estresse oxidativo. Neste sentido, acreditamos que a atenuação da disfunção autonômica (precocemente observada neste modelo de DM) pelo treinamento físico, independente do tipo, possa induzir alterações favoráveis (e dependentes do tipo de treino) no sistema renina angiotensina e em mediadores inflamatórios, reduzindo o estresse oxidativo em tecidos importantes para a regulação cardiovascular / The aim of this study was to evaluate the role of aerobic, resistance or combined (aerobic + resistance) exercise training in the development of type 2 diabetes, analyzing mechanisms associated with diabetes complications in female mice with deficiency in leptin production (ob/ob). Female mice, initially with 4 weeks of age, were divided into 6 groups: ob/ob sedentary with 4 weeks of life (OS-4), sedentary wild type (SS) or ob/ob sedentary (OS-12) followed until 12 week life, ob/ob+aerobic training (OA), ob/ob+resistance training (OR) and ob/ob+combined training (OC). The trained groups were submitted to eight weeks of dynamic aerobic exercise training on a treadmill (50-60% of maximum stress test speed) or resistance exercise on a ladder (40-60% of the maximum load) or an association of these two trainings (combined). Body weight; glucose, triglycerides, and total blood cholesterol; blood pressure (BP) and heart rate (HR); baroreflex sensitivity (BRS); cardiovascular autonomic modulation; inflammatory and hormonal markers; and oxidative stress parameters were evaluated. Obese animals (OS-12) showed increased body and fat weight, blood glucose, triglyceride and glucose intolerance when compared to wild type animals (SS). Additionally, OS-12 group showed decreased capacity in aerobic and strength exercise tests. We did not observe differences between the SS and the OS-12 groups regarding BP and HR, however the OS-12 group showed reduced heart rate variability (HRV) (33 ± 4ms2) and baroreflex sensitivity when compared to the SS group (178 ± 19 ms2). The OS-12 group showed increased angiotensin 2 in kidney and heart tissues, decreased adiponectin and increased inflammatory cytokines in adipose tissue and in the spleen in relation to the SS group. Moreover, obese animals presented increased protein oxidation and lipid peroxidation and decreased antioxidant enzymes in kidney and heart tissues when compared to SS group. The comparison between the OS-4 and the OS-12 groups showed increased body and fat weight, blood glucose, glucose intolerance and oxidative stress in OS-12 compared to OS-4 group. The decrease in HRV and in BRS were observed in OS-4 and OS-12 groups. On the other hand, exercise training decreased weight gain and reduced blood glucose and glucose intolerance in all three trained groups compared to OS-12 group. Aerobic (61 ± 8ms2 and 6 ± 4 mmHg2) and resistance exercise training (66 ± 16ms2 and 6 ± 1.4mmHg2) were efficient in increasing the HRV and decrease the low frequency band of BP (vascular sympathetic modulation) as compared to OS-12 group; however OC group (43 ± 7ms2 and 8 ± 0.9mmHg2) was similar to OS-12 group (10±1.1mmHg2) and to OA and OR groups. In addition, the three types of exercsie training improved BRS. The three types of training reduced levels of angiotensin 2 and increased levels of angiotensin 1-7 in adipose tissue, kidney and heart. The aerobic exercise was more efficient in improving inflammatory profile in adipose tissue and spleen, since OA and OC groups showed an increase in adiponectin and only the OC group showed a decrease in IL-6 and PAI-1 in relation to the group OS-12. Regarding oxidative stress, the three trained groups showed a decrease in damage markers. In conclusion, our findings support the development of metabolic dysfunction during lifespan in ob/ob mice. Interestingly, 4 weeks old ob/ob mice showed a significant reduction in HRV parameters. This autonomic imbalance could be occurring not only in the heart, but in other tissues, such as the spleen and the adipose tissue, promoting the release of inflammatory cytokines. These cytokines could induce long-term target organ damage, as observed in this study in heart and kidney by increased oxidative stress. The trained groups, regardless of the type of training, showed improved metabolic and cardiovascular autonomic regulation, which were accompanied by favorable changes in the renin-angiotensin system, inflammatory mediators and oxidative stress profile. In this sense, we believe that the attenuation of autonomic dysfunction (early observed in this model of DM) by exercise training, regardless of the type of training, can induce positive changes (dependent on the type of exercise training) on the renin angiotensin system and inflammatory mediators, reducing oxidative stress in important tissues for cardiovascular regulation

O objetivo do presente estudo foi avaliar o papel do treinamento físico aeróbio, resistido ou combinado (aeróbio+resistido) no desenvolvimento do diabetes tipo 2 analisando mecanismos associados às complicações no diabetes em camundongos fêmeas com deficiência na produção leptina (ob/ob). Para tanto, foram utilizadas camundongos fêmeas, inicialmente com 4 semanas de idade, divididas em 6 grupos: ob/ob sedentárias com 4 semanas de vida (OS-4), selvagens sedentárias (SS) ou ob/ob sedentárias (OS-12) acompanhadas até a 12ª semana de vida, ob/ob treinamento aeróbio (OA), ob/ob treinamento resistido (OR) e ob/ob treinamento combinado (OC). Os grupos treinados foram submetidos a 8 semanas de treinamento físico dinâmico aeróbio em esteira (50 a 60% da velocidade máxima do teste de esforço) ou resistido em escada (4060% da carga máxima) ou a associação dos dois treinos (combinado). Foram avaliados: peso corporal; glicose, triglicérides e colesterol total sanguíneos; pressão arterial (PA) e frequência cardíaca (FC); sensibilidade barorreflexa (SBR); modulação autonômica cardiovascular; marcadores inflamatórios e hormonais; e parâmetros de estresse oxidativo. Os animais obesos (OS-12) apresentaram aumento de peso corporal, tecido adiposo, de glicemia, de triglicérides e de intolerância à glicose quando comparado aos animais selvagens (SS). Adicionalmente, o grupo OS-12 apresentou piores resultados nos testes aeróbio e de força. Não observamos diferenças entre os grupos SS e OS-12 em relação a PA e FC, porém o grupo OS-12 apresentou diminuição da variabilidade da frequência cardíaca (VFC) (33 ± 4ms2) e da sensibilidade barorreflexa em relação ao grupo SS (VFC: 178 ± 19 ms2). O grupo OS-12 apresentou aumento de angiotensina 2 nos tecidos renal e cardíaco, diminuição da adiponectina e aumento de citocinas inflamatórias no tecido adiposo e no baço em relação ao grupo SS. Somado a isso, os animais obesos apresentam maior dano a proteínas e lipoperoxidação e diminuição das enzimas antioxidantes em tecido renal e cardíaco em relação ao grupo SS. A comparação entre os grupos OS-4 e OS-12 evidenciou aumento de peso corporal, tecido adiposo, glicemia, intolerância à glicose e de parâmetros de estresse oxidativo no grupo OS-12 em relação ao grupo OS-4. A redução na VFC e na SBR foi observada no grupo OS-4 e no grupo OS-12. O treinamento físico por sua vez, diminuiu o ganho de peso e reduziu a glicemia e a intolerância à glicose nos três grupos treinados em comparação ao grupo OS-12. O treinamento físico aeróbio (61 ± 8ms2 e 6 ± 4 mmHg2) e resistido (66 ± 16ms2 e 6 ± 1,4mmHg2) foram eficientes em aumentar a VFC e diminuir a banda de baixa frequência da PA (simpático vascular) em relação ao grupo OS-12, porém o grupo OC (43 ± 7ms2 e 8 ± 0,9mmHg2) foi semelhante ao grupo OS-12 (10 ± 1,1mmHg2) e aos grupos OA e OR. Além disso, as três modalidades melhoraram a SBR. Os três tipos de treinamento reduziram os níveis de angiotensina 2 e aumentaram os níveis de angiotensina 1-7 em tecido adiposo, rim e coração. O treinamento físico aeróbio foi mais eficiente em melhorar o perfil inflamatório em tecido adiposo e no baço, uma vez que os grupos OA e OC apresentaram aumento de adiponectina e apenas o grupo OC apresentou diminuição de IL-6 e PAI-1 em relação ao grupo OS-12. Em relação ao estresse oxidativo, os três grupos treinados apresentaram diminuição de marcadores de lesão. Concluindo, nossos achados confirmam o desenvolvimento de disfunção metabólica ao longo da vida de camundongos ob/ob. É interessante notar que com 4 semanas de vida camundongos ob/ob apresentaram uma expressiva redução dos parâmetros da VFC. Este desbalanço autonômico, poderiam estar ocorrendo não só no coração, mas para outros tecidos, como o baço e o tecido adiposo, favorecendo a liberação de citocinas inflamatórias que poderiam induzir a longo prazo lesão de órgão alvo, como observado no presente estudo em coração e rins, por aumento de estresse oxidativo. Os grupos treinados, independente da modalidade, apresentaram melhora metabólica e na regulação autonômica cardiovascular, a qual foi acompanhada de alterações favoráveis no sistema renina-angiotensina, em mediadores inflamatórios e no perfil de estresse oxidativo. Neste sentido, acreditamos que a atenuação da disfunção autonômica (precocemente observada neste modelo de DM) pelo treinamento físico, independente do tipo, possa induzir alterações favoráveis (e dependentes do tipo de treino) no sistema renina angiotensina e em mediadores inflamatórios, reduzindo o estresse oxidativo em tecidos importantes para a regulação cardiovascular / The aim of this study was to evaluate the role of aerobic, resistance or combined (aerobic + resistance) exercise training in the development of type 2 diabetes, analyzing mechanisms associated with diabetes complications in female mice with deficiency in leptin production (ob/ob). Female mice, initially with 4 weeks of age, were divided into 6 groups: ob/ob sedentary with 4 weeks of life (OS-4), sedentary wild type (SS) or ob/ob sedentary (OS-12) followed until 12 week life, ob/ob+aerobic training (OA), ob/ob+resistance training (OR) and ob/ob+combined training (OC). The trained groups were submitted to eight weeks of dynamic aerobic exercise training on a treadmill (50-60% of maximum stress test speed) or resistance exercise on a ladder (40-60% of the maximum load) or an association of these two trainings (combined). Body weight; glucose, triglycerides, and total blood cholesterol; blood pressure (BP) and heart rate (HR); baroreflex sensitivity (BRS); cardiovascular autonomic modulation; inflammatory and hormonal markers; and oxidative stress parameters were evaluated. Obese animals (OS-12) showed increased body and fat weight, blood glucose, triglyceride and glucose intolerance when compared to wild type animals (SS). Additionally, OS-12 group showed decreased capacity in aerobic and strength exercise tests. We did not observe differences between the SS and the OS-12 groups regarding BP and HR, however the OS-12 group showed reduced heart rate variability (HRV) (33 ± 4ms2) and baroreflex sensitivity when compared to the SS group (178 ± 19 ms2). The OS-12 group showed increased angiotensin 2 in kidney and heart tissues, decreased adiponectin and increased inflammatory cytokines in adipose tissue and in the spleen in relation to the SS group. Moreover, obese animals presented increased protein oxidation and lipid peroxidation and decreased antioxidant enzymes in kidney and heart tissues when compared to SS group. The comparison between the OS-4 and the OS-12 groups showed increased body and fat weight, blood glucose, glucose intolerance and oxidative stress in OS-12 compared to OS-4 group. The decrease in HRV and in BRS were observed in OS-4 and OS-12 groups. On the other hand, exercise training decreased weight gain and reduced blood glucose and glucose intolerance in all three trained groups compared to OS-12 group. Aerobic (61 ± 8ms2 and 6 ± 4 mmHg2) and resistance exercise training (66 ± 16ms2 and 6 ± 1.4mmHg2) were efficient in increasing the HRV and decrease the low frequency band of BP (vascular sympathetic modulation) as compared to OS-12 group; however OC group (43 ± 7ms2 and 8 ± 0.9mmHg2) was similar to OS-12 group (10±1.1mmHg2) and to OA and OR groups. In addition, the three types of exercsie training improved BRS. The three types of training reduced levels of angiotensin 2 and increased levels of angiotensin 1-7 in adipose tissue, kidney and heart. The aerobic exercise was more efficient in improving inflammatory profile in adipose tissue and spleen, since OA and OC groups showed an increase in adiponectin and only the OC group showed a decrease in IL-6 and PAI-1 in relation to the group OS-12. Regarding oxidative stress, the three trained groups showed a decrease in damage markers. In conclusion, our findings support the development of metabolic dysfunction during lifespan in ob/ob mice. Interestingly, 4 weeks old ob/ob mice showed a significant reduction in HRV parameters. This autonomic imbalance could be occurring not only in the heart, but in other tissues, such as the spleen and the adipose tissue, promoting the release of inflammatory cytokines. These cytokines could induce long-term target organ damage, as observed in this study in heart and kidney by increased oxidative stress. The trained groups, regardless of the type of training, showed improved metabolic and cardiovascular autonomic regulation, which were accompanied by favorable changes in the renin-angiotensin system, inflammatory mediators and oxidative stress profile. In this sense, we believe that the attenuation of autonomic dysfunction (early observed in this model of DM) by exercise training, regardless of the type of training, can induce positive changes (dependent on the type of exercise training) on the renin angiotensin system and inflammatory mediators, reducing oxidative stress in important tissues for cardiovascular regulation

Purpose: This study will identify that patients with chronic neck pain have an altered autonomic functioning compared to a control group, and that manipulation might directly influence the autonomic nervous system as measured by using a fully automated pupillometry system. Subjects: 100 chronic pain patients and 50 control subjects participated in this study to achieve a power of 0.80, effect size of 0.5, and a type I error rate of 0.05 for two-tailed hypothesis testing. Method: A quasi-experimental design was be used. The ANOVA and Chi square test were used to establish homogeneity of baseline characteristics. The Mann-Whitney U test was performed to compare the pre-intervention pupil diameter amongst the groups. The Friedman's test was used to determine the pupil diameter change during the three measurements. The Wilcoxen Signed-ranks test was used to analyze the difference in pupil size between the pre- and post-intervention measures and to determine if there was a difference in pupil diameter between the two groups undergoing the thoracic manipulation. The Kruskal-Wallis test was used to compare the pupil diameter change to the presence of joint sounds. And the Fisher's Exact test was used to determine the relationship between the number of pops and the VAS change score > 13mm. Results: This study demonstrated that the chronic pain group had a statistically significant smaller pupil diameter than the healthy control group (P=0.022). Manipulation resulted in a relative increase in pupil diameter following the manipulation, however this was not statistical significant. There was a statistical significant decrease in pupil diameter in the placebo group (pConclusions: It appears that a T3-T4 manipulation results in a relative non-specific increase in sympathetic activity. Recommendations: Manipulation may be used by physical therapists to affect the autonomic nervous system. Visual pupil assessment may become part of the evaluation process to identify patients that might present with autonomic dysfunction and to determine the effect of treatment modalities.

No description available.

Chronic refractory gastrointestinal (GI) motility disorders are a significant burden on the healthcare system, acting as a large public health issue with significant impact on the quality of life in both the pediatric and adult population. Control systems of gastrointestinal motility are complex and involve coordination of smooth muscle contraction and relaxation, which the autonomic nervous system is largely responsible for. Gaps in the diagnosis process, such as overlooking autonomic function, has left patients with diminished quality of life and limited treatment options. Many patients in the clinic have experienced injury within the spinal cord and we hypothesized that GI symptoms might be related to spinal injury causing disruption of sensory and/or motor nerves of the autonomic nervous system. Our objective became to better understand the specific location and nature of spinal injuries and GI symptoms, as completed through the development of a self-report questionnaire. Main findings suggest symptoms indicative of T3-T9 and T10-L2 spinal pathology. COVID-19 did not allow for in-clinic neuromodulation with autonomic assessments, resulting in experiments remotely assessing at-home neuromodulation treatment for GI symptoms with suspected spinal autonomic dysfunction. At-home neuromodulation was not suitable for many patients, but those who were able to manage it showed highly promising results. After years of suffering, transcutaneous electrical nerve stimulation alleviated symptoms, particularly postprandial abdominal pain, constipation, vomiting and nausea. I discuss what we learned to set us up for successful at-home treatment, and we will use all information to design randomized controlled trials to prove the benefit of TENS. The present work offers significant information on the relationship of thoracolumbar spinal pathology and complex GI symptoms, which is now used in the clinic in the diagnosis process of GI dysmotility. In addition, we have learned how to conduct at-home treatment using TENS, which allows us to execute future studies. / Thesis / Master of Science (MSc) / There are gaps in the diagnosis process of complex gastrointestinal (GI) dysmotility disorders, including lack of testing of autonomic function, leaving patients suffering with diminished quality of life with unsuccessful treatment attempts. As many patients also experience injury or conditions of the spine, I have hypothesized that GI symptoms may be related to spinal injury-induced dysfunction of the autonomic nervous system. Experimental models aim to understand the location and nature of spinal pathology with GI symptoms for future diagnoses, as well as potential treatment options such as neuromodulation. Findings of this thesis suggest symptoms indicative of particular thoracolumbar spinal pathology and promising results of transcutaneous electrical nerve stimulation (TENS) to alleviate GI symptoms, including T3-T9 and T10-L2 spinal pathology-related postprandial abdominal pain, constipation, nausea, and vomiting. This work offers information for the diagnostic process of GI dysmotility and the future design of clinical trials of neuromodulation therapies.

There are approximately 6000 to 8000 rare diseases, each with a prevalence of less than 1 / 10 000, but in aggregate affecting 6 to 8% of the population. It is important to evaluate disease development and progression to know the natural course of any disease. This information can be utilized in diagnostics and in assessing effects of therapeutic interventions as they become available. This thesis describes the natural clinical history and evolution of imaging findings of two rare diseases over approximately two decades. Papers I, II and III present clinical, magnetic resonance imaging (MRI), magnetic resonance spectroscopy (MRS) and 18F-fluorodeoxyglucose positron emission tomography (FDG-PET) findings in LMNB1-related autosomal dominant leukodystrophy (ADLD). MRI was found to be very sensitive in finding pathology in patients with LMNB1-related ADLD, even before the onset of clinical symptoms. However, even patients with widespread MRI changes can have a relatively mild symptomatology and present only slight disturbances in metabolic examinations such as MRS and FDG-PET. This is compatible with relatively intact axons, even as myelin impairment is widespread. Paper IV presents clinical and MRI findings in the brain and musculature in SIL1-positive Marinesco-Sjögren syndrome (MSS), and describes a new, mild phenotype of the disease with no intellectual disabilities and only slight motor disabilities. With a 19-year-long radiological follow-up, a slow progressive atrophic process in the cerebellum and brainstem could be demonstrated. MRI of the musculature shows early involvement of the quadriceps and gastrocnemii but not the tibialis anterior, progressing to widespread atrophy in the back and upper and lower limbs at the age of 20 years. In the mildest phenotype, the most severely affected muscles were the m gluteus maximus, m sartorius, m peroneus longus, and the lateral head of the m gastrocnemius.

Spinal cord injury (SCI) remains a significant cause for morbidity and mortality in North America. Bladder dysfunction following SCI is very common and could lead to severe complications including renal failure and autonomic dysreflexia (AD). AD involves life threatening episodes of hypertension in patients with SCI above T6 level. Current management protocols for AD are symptomatic and usually ineffective. Botulinum toxin-A (BTX-A), has been successfully used recently in SCI patients because it reduces the detrusor contractility via inhibiting acetylcholine release from efferent nerve endings. Recent evidence, however, suggests a sensory involvement via modulation of sensory neuropeptides, neurotransmitters, and receptors. It is still, however, unclear whether BTX-A can affect putative spinal neurons involved in AD. In this study we demonstrated that intravesical BTX-A treatment has blocked AD in rats with T4-SCI, and also provided a novel mechanism for the control of autonomic dysreflexia via a minimally invasive treatment modality.

Spinal cord injury (SCI) remains a significant cause for morbidity and mortality in North America. Bladder dysfunction following SCI is very common and could lead to severe complications including renal failure and autonomic dysreflexia (AD). AD involves life threatening episodes of hypertension in patients with SCI above T6 level. Current management protocols for AD are symptomatic and usually ineffective. Botulinum toxin-A (BTX-A), has been successfully used recently in SCI patients because it reduces the detrusor contractility via inhibiting acetylcholine release from efferent nerve endings. Recent evidence, however, suggests a sensory involvement via modulation of sensory neuropeptides, neurotransmitters, and receptors. It is still, however, unclear whether BTX-A can affect putative spinal neurons involved in AD. In this study we demonstrated that intravesical BTX-A treatment has blocked AD in rats with T4-SCI, and also provided a novel mechanism for the control of autonomic dysreflexia via a minimally invasive treatment modality.

Hypertension is associated with marked cardiac sympathetic over-activity and end organ hyper-responsiveness. The sympathetic dysfunction is caused by aberrant calcium (Ca²⁺) handling resulting in enhanced neurotransmission. However, it remains unclear whether the sympathetic neuron or the myocytes is the primary driver behind the initiation and maintenance of the autonomic phenotype. The work in this thesis characterises the Ca²⁺ dysfunction and regulation at the membrane level. Further, it employs physiologically coupled sympathetic neurons and ventricular myocytes to determine the cellular driver of cardiac dysautonomia in the pro-hypertensive state. <b>Chapter 1</b> provides a general overview of the field of autonomic hypertension with a specific focus on the sympathetic control of cardiac excitability. In particular, the role of Ca²⁺ and cyclic nucleotides in the facilitation of neurotransmission are explored. <b>Chapter 2</b> details the methods used in this thesis. It provides rationale for the approaches taken to record membrane Ca2+ currents, cyclic adenosine monophosphate (cAMP) levels and cAMP-activated protein kinase (PKA) activity, and the development and uses of a co-culture of coupled sympathetic neurons and ventricular myocytes. <b>Chapter 3</b> describes the successful development of an effective voltage clamp method to isolate whole cell Ca²⁺ currents in sympathetic neurons. It details the issue of space clamp problem when using this technique on peripheral neurons and provides experimental guidance on how to quantify and limit theses issues. <b>Chapter 4</b> identifies that the pro-hypertensive four-week old neurons from the spontaneously hypertensive rat (SHR) have significantly larger whole cell Ca²⁺ currents when compared to normotensive (Wistar Kyoto-WKY) neurons, that are largely N-type in nature. Restoring the cGMP cyclic nucleotide dysfunction seen in these cells, rescues the ion channel phenotype and bring the Ca²⁺ down to levels seen in the normotensive WKY neuron. Further, it identifies that phosphodiesterase (PDE) 2A inhibition differentially affects the currents in the WKY and SHR, further supporting the notion of PDE2A dominance. <b>Chapter 5</b> identifies the presence and functional relevance of cGMP cross-talk with the cAMP-PKA pathway in sympathetic neurons. This cross talk is significantly altered in the pro-hypertensive state, via the differential involvement of PDEs. It functionally identifies the presence of PDE3 and PDE2A and provides further evidence that these enzymes could be dysregulated in pro-hypertensive neurons. <b>Chapter 6</b> describes the use of a co-culture model of ventricular myocytes and sympathetic neurons. Physiological stimulation of the sympathetic neuron with nicotine whilst monitoring cAMP levels in the myocytes confirms that the cellular phenotypes seen in the individual cells are functionally present in the co-culture. Using cross-cultures, it identifies the neuron as the principal driver behind the cardiac sympathetic responses observed in pro-hypertension. The results provide evidence for a dominant role played by the neuron in driving the adrenergic phenotype seen in cardiovascular disease and highlights the potential of using healthy neurons to turn down the gain of neurotransmission, akin to a smart pre-synaptic &beta;-blocker. <b>Chapter 7</b> forms the concluding discussion that summarises the main findings of this thesis and attempt to place it in a clinical context, and highlights avenues of further research. In particular, the possibility of using a cell therapeutic approach to treat sympathetic hyperactivity.

Submitted by isabela.moljf@hotmail.com (isabela.moljf@hotmail.com) on 2017-05-18T15:29:32Z No. of bitstreams: 1 carlosalbertodeoliveira.pdf: 972558 bytes, checksum: b1b7c46020558acfe2bd3a27284c43ff (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2017-05-18T15:44:56Z (GMT) No. of bitstreams: 1 carlosalbertodeoliveira.pdf: 972558 bytes, checksum: b1b7c46020558acfe2bd3a27284c43ff (MD5) / Made available in DSpace on 2017-05-18T15:44:56Z (GMT). No. of bitstreams: 1 carlosalbertodeoliveira.pdf: 972558 bytes, checksum: b1b7c46020558acfe2bd3a27284c43ff (MD5) Previous issue date: 2010-07-30 / As doenças cardiovasculares são as principais causas de morbidade e mortalidade em indivíduos com doença renal crônica (DRC). Esses achados se devem à presença de múltiplos fatores de risco para aterosclerose como o diabetes mellitus, a dislipidemia, a hipertensão arterial e a hipertrofia ventricular esquerda, bem como de fatores não tradicionais como mediadores inflamatórios, anemia e calcificação vascular. Além desses, a disfunção do sistema nervoso autônomo (SNA) tem sido associada à maior prevalência de arritmias cardíacas e de morte súbita nessa população. Objetivo: no presente estudo, avaliou-se a variabilidade da freqüência cardíaca (VFC) através da análise espectral pelo sistema holter, em repouso e sob estresse postural passivo em pacientes renais crônicos sob tratamento conservador. Casuística e métodos: foram avaliados 32 pacientes renais crônicos em tratamento conservador nos estágios 3, 4 e 5 (grupo DRC) e 14 voluntários saudáveis (grupo CON). Todos os indivíduos foram submetidos à avaliação cardiológica, com realização de eletrocardiograma e de ecodopplercardiograma, seguidos da análise da VFC. Para tanto, foi feito o registro contínuo da freqüência cardíaca através do sistema holter, durante dois períodos de 20 minutos, em decúbito dorsal (período pré-inclinado) e em estresse postural passivo a 70º (estresse ortostático). A VFC foi obtida através da análise espectral das variáveis de baixa freqüência normalizada (LFnu), de alta freqüência normalizada (HFnu), indicativas da modulação simpática e parassimpática, respectivamente, além da razão LFnu/HFnu, indicativa do balanço simpático-vagal.Análise estatística: as variáveis foram testadas usando-se o teste de normalidade de Kolmogorov-Smirnov, que mostrou condição de não-normalidade dos dados obtidos pela análise espectral. Desse modo, as hipóteses de igualdade dos valores foram analisadas pelos testes não-paramétricos de Mann-Whitney, Wilcoxon e Kruskal-Wallis. Para variáveis normais utilizou-se o teste T Student e ANOVA. Para avaliar a correlação entre a filtração glomerular estimada e a razão LFnu/HFnu em pacientes com DRC, usou-se o coeficiente de correlação linear de Spearman. Os resultados foram expressos em média (X) ± desvio-padrão (DP). Um valor de p menor ou igual a 0,05 foi considerado significativo. Resultados: no período pré-inclinado não foram observadas diferenças significativas das variáveis analisadas entre os grupos DRC e CON. No entanto, após a inclinação, a variável LFnu foi menor (73±13,0 versus 84,5±8, p<0,05), a variável HFnu foi maior (26,5±13,0 versus 15,4±8,1, p<0,05), enquanto a razão LFnu/HFnu foi significativamente menor (3,3±1,3 versus 7,9±5,6, p<0,05) no grupo DRC comparado ao grupo CON. A comparação entre os pacientes com DRC nos estágios 3,4 e 5, no período pré-inclinado não mostrou diferenças significativas das variáveis LFnu, HFnu e razão LFnu/HFnu. Entretanto, durante o período inclinado, os pacientes do estágio 5 apresentaram LFnu menor, HFnu maior e razão LFnu/HFnu menor que pacientes no estágio 3 (66,9±12,9 versus 78,5±5,2; 33,1±13,0 versus 21,5±5,2; e 2,4±1,2 versus 3,9±1,2; respectivamente) (p<0.05). Conclusão: Pacientes com DRC em tratamento conservador apresentam disautonomia cardíaca que piora com a redução do ritmo de filtração glomerular. / Cardiovascular diseases are the leading cause of morbidity and mortality in subjects with chronic kidney disease (CKD). These findings are related to the presence of multiple risk factors for atherosclerosis such as diabetes, dislipidemia, hypertension, and left ventricular hypertrophy, as well as to non traditional risk factors like inflammatory mediators, anemia, and vascular calcification. Furthermore, autonomic nervous system dysfunction has been associated to higher prevalence of cardiac arrhythmias and sudden death in this population. Objectives: In the present study, the heart rate variability (HRV) was evaluated through the spectral analysis using a holter monitoring, both in the supine position and upon passive postural stress in patients with CKD on conservative treatment. Patients and Methods: We evaluated 32 patients with CKD on conservative treatment stages 3, 4 and 5 (CRD group) and 14 healthy volunteers (CON group). All subjects underwent cardiological evaluation including electrocardiogram, and doppler echocardiogram, followed by the analysis of HRV. Heart rate variability was performed by continuous recording of heart rate by holter monitoring system during two periods of 20 minutes in supine position and passive postural tilt of 70° (orthostatic stress). HRV was obtained by spectral analysis of the variables normalized low frequency (LFnu), normalized high frequency (HFnu), indicative of sympathetic and parasympathetic modulation, respectively, as well as the LHnu/HFnu ratio, indicative of sympathovagal balance. Variables were tested for normality using the Kolmogorov-Smirnov test, which showed the condition of non-normality of the data obtained by spectral analysis. Thus, the hypotheses of equality of the values were analyzed with the Mann-Whitney, Wilcoxon, and Kruskal-Wallis non-parametric tests. For normal variables we used unpaired Student´s T test and ANOVA. To assess the correlation between the eGFR and the LFnu/HFnu ratio in patients with CKD, we used Spearman linear correlation coefficient. Results were expressed as the mean (X) ± standard deviation (SD). A p value less than or equal to 0.05 was considered statistically significant. Results: During the pre-inclined period no significant differences among variables in the groups CKD and CON were observed. However, during the inclined period, the variable LFnu was lower (73 ± 13.0 versus 84.5 ± 8, p <0.05), the variable HFnu was higher (26.5 ± 13.0 versus 15.4 ± 8.1, p <0.05), while the LFnu/HFnu ratio was lower (3.3 ± 1.3 versus 7.9 ± 5.6, p <0.05) in the CKD group compared to the CON group. The comparison between patients with CKD in stages 3, 4 and 5, in the pre-inclined period did not show significant differences in the variables LFnu, HFnu and LFnu/HFnu ratio. However, during the inclined period, stage 5 patients showed lower LFnu, higher HFnu and LFnu/HFnu ratio lower than in stage 3 patients (66.9 ± 12.9 versus 78.5 ± 5.2, 33.1 ± 13.0 versus 21.5 ± 5.2 and 2.4 ± 1.2 versus 3.9 ± 1.2, respectively) (p <0.005). Conclusion: Patients with CKD present depressed cardiac autonomic modulation that worsens with decreasing glomerular filtration rate.