Progressive degeneration in the central nervous system (CNS) of Huntington’s disease (HD) patients is a relentless debilitating process, resulting from the inheritance of a single gene mutation. With limited knowledge of the underlying pathological molecular mechanisms, pharmaceutical intervention has to-date not provided any effective clinical treatment strategies to attenuate or compensate the neuronal cell death. Attention has therefore turned to biotherapeutic molecules and novel treatment approaches to promote restoration and protection of selectively vulnerable populations of neurons in the HD brain. Rapid advances in vectorology and gene-based medicine over the past decade have opened the way for safe and efficient delivery of biotherapeutics to the CNS. With numerous factors known to regulate the development, plasticity and maintenance of the mammalian nervous system many proteins have emerged as potential therapeutic agents to alleviate HD progression. This investigative study utilised gene delivery vectors derived from the non-pathogenic adeno-associated virus (AAV) to direct high-level expression of brain-derived neurotrophic factor (BDNF), glial cell-line derived neurotrophic factor (GDNF), Bcl-xL or X-linked inhibitor of apoptosis protein (XIAP) within the rodent striatum. Maintenance of the basal ganglia and functional behaviour deficits were assessed following excitotoxic insult of the striatum by quinolinic acid (QA), a neurotoxic model of HD pathology. Enhanced striatal expression of BDNF prior to QA-induced lesioning provided maintenance of the striosome-matrix organisation of the striatum, attenuating impairments of sensorimotor behaviour with a 36-38% increase in the maintenance of DARPP-32 / krox-24 expressing striatal neurons, reduced striatal atrophy and increased maintenance of striatonigral projections. Higher levels of BDNF however induced seizures and weight-loss highlighting the need to provide regulatable control over biotherapeutic protein expression. Continuous high-expression of BDNF or GDNF resulted in a downregulation of intracellular signal mediating proteins including DARPP-32, with AAV-GDNF not found to enhance the overall maintenance of striatal neurons. Neither of the anti-apoptotic factors provided significant protection of transduced striatal neurons but tended towards ameliorating QA-induced behavioural deficits, displaying behaviour – pathology correlations with the survival of parvalbumin-expressing neurons in the globus pallidus. The results of this thesis suggest BDNF as a promising putative biotherapeutic for HD, but emphasises the requirement to control expression following gene delivery, and for further elucidation of the physiological impact that enhanced expression of endogenous factors has on the host cells. Additionally the maintenance of neural networks beyond the caudate-putamen will be vital to ensuring efficient clinical outcomes for HD. / Auckland Medical Research Foundation. Foundation for Research, Science and Technology. The University of Auckland.

Progressive degeneration in the central nervous system (CNS) of Huntington’s disease (HD) patients is a relentless debilitating process, resulting from the inheritance of a single gene mutation. With limited knowledge of the underlying pathological molecular mechanisms, pharmaceutical intervention has to-date not provided any effective clinical treatment strategies to attenuate or compensate the neuronal cell death. Attention has therefore turned to biotherapeutic molecules and novel treatment approaches to promote restoration and protection of selectively vulnerable populations of neurons in the HD brain. Rapid advances in vectorology and gene-based medicine over the past decade have opened the way for safe and efficient delivery of biotherapeutics to the CNS. With numerous factors known to regulate the development, plasticity and maintenance of the mammalian nervous system many proteins have emerged as potential therapeutic agents to alleviate HD progression. This investigative study utilised gene delivery vectors derived from the non-pathogenic adeno-associated virus (AAV) to direct high-level expression of brain-derived neurotrophic factor (BDNF), glial cell-line derived neurotrophic factor (GDNF), Bcl-xL or X-linked inhibitor of apoptosis protein (XIAP) within the rodent striatum. Maintenance of the basal ganglia and functional behaviour deficits were assessed following excitotoxic insult of the striatum by quinolinic acid (QA), a neurotoxic model of HD pathology. Enhanced striatal expression of BDNF prior to QA-induced lesioning provided maintenance of the striosome-matrix organisation of the striatum, attenuating impairments of sensorimotor behaviour with a 36-38% increase in the maintenance of DARPP-32 / krox-24 expressing striatal neurons, reduced striatal atrophy and increased maintenance of striatonigral projections. Higher levels of BDNF however induced seizures and weight-loss highlighting the need to provide regulatable control over biotherapeutic protein expression. Continuous high-expression of BDNF or GDNF resulted in a downregulation of intracellular signal mediating proteins including DARPP-32, with AAV-GDNF not found to enhance the overall maintenance of striatal neurons. Neither of the anti-apoptotic factors provided significant protection of transduced striatal neurons but tended towards ameliorating QA-induced behavioural deficits, displaying behaviour – pathology correlations with the survival of parvalbumin-expressing neurons in the globus pallidus. The results of this thesis suggest BDNF as a promising putative biotherapeutic for HD, but emphasises the requirement to control expression following gene delivery, and for further elucidation of the physiological impact that enhanced expression of endogenous factors has on the host cells. Additionally the maintenance of neural networks beyond the caudate-putamen will be vital to ensuring efficient clinical outcomes for HD. / Auckland Medical Research Foundation. Foundation for Research, Science and Technology. The University of Auckland.

Progressive degeneration in the central nervous system (CNS) of Huntington’s disease (HD) patients is a relentless debilitating process, resulting from the inheritance of a single gene mutation. With limited knowledge of the underlying pathological molecular mechanisms, pharmaceutical intervention has to-date not provided any effective clinical treatment strategies to attenuate or compensate the neuronal cell death. Attention has therefore turned to biotherapeutic molecules and novel treatment approaches to promote restoration and protection of selectively vulnerable populations of neurons in the HD brain. Rapid advances in vectorology and gene-based medicine over the past decade have opened the way for safe and efficient delivery of biotherapeutics to the CNS. With numerous factors known to regulate the development, plasticity and maintenance of the mammalian nervous system many proteins have emerged as potential therapeutic agents to alleviate HD progression. This investigative study utilised gene delivery vectors derived from the non-pathogenic adeno-associated virus (AAV) to direct high-level expression of brain-derived neurotrophic factor (BDNF), glial cell-line derived neurotrophic factor (GDNF), Bcl-xL or X-linked inhibitor of apoptosis protein (XIAP) within the rodent striatum. Maintenance of the basal ganglia and functional behaviour deficits were assessed following excitotoxic insult of the striatum by quinolinic acid (QA), a neurotoxic model of HD pathology. Enhanced striatal expression of BDNF prior to QA-induced lesioning provided maintenance of the striosome-matrix organisation of the striatum, attenuating impairments of sensorimotor behaviour with a 36-38% increase in the maintenance of DARPP-32 / krox-24 expressing striatal neurons, reduced striatal atrophy and increased maintenance of striatonigral projections. Higher levels of BDNF however induced seizures and weight-loss highlighting the need to provide regulatable control over biotherapeutic protein expression. Continuous high-expression of BDNF or GDNF resulted in a downregulation of intracellular signal mediating proteins including DARPP-32, with AAV-GDNF not found to enhance the overall maintenance of striatal neurons. Neither of the anti-apoptotic factors provided significant protection of transduced striatal neurons but tended towards ameliorating QA-induced behavioural deficits, displaying behaviour – pathology correlations with the survival of parvalbumin-expressing neurons in the globus pallidus. The results of this thesis suggest BDNF as a promising putative biotherapeutic for HD, but emphasises the requirement to control expression following gene delivery, and for further elucidation of the physiological impact that enhanced expression of endogenous factors has on the host cells. Additionally the maintenance of neural networks beyond the caudate-putamen will be vital to ensuring efficient clinical outcomes for HD. / Auckland Medical Research Foundation. Foundation for Research, Science and Technology. The University of Auckland.

Progressive degeneration in the central nervous system (CNS) of Huntington’s disease (HD) patients is a relentless debilitating process, resulting from the inheritance of a single gene mutation. With limited knowledge of the underlying pathological molecular mechanisms, pharmaceutical intervention has to-date not provided any effective clinical treatment strategies to attenuate or compensate the neuronal cell death. Attention has therefore turned to biotherapeutic molecules and novel treatment approaches to promote restoration and protection of selectively vulnerable populations of neurons in the HD brain. Rapid advances in vectorology and gene-based medicine over the past decade have opened the way for safe and efficient delivery of biotherapeutics to the CNS. With numerous factors known to regulate the development, plasticity and maintenance of the mammalian nervous system many proteins have emerged as potential therapeutic agents to alleviate HD progression. This investigative study utilised gene delivery vectors derived from the non-pathogenic adeno-associated virus (AAV) to direct high-level expression of brain-derived neurotrophic factor (BDNF), glial cell-line derived neurotrophic factor (GDNF), Bcl-xL or X-linked inhibitor of apoptosis protein (XIAP) within the rodent striatum. Maintenance of the basal ganglia and functional behaviour deficits were assessed following excitotoxic insult of the striatum by quinolinic acid (QA), a neurotoxic model of HD pathology. Enhanced striatal expression of BDNF prior to QA-induced lesioning provided maintenance of the striosome-matrix organisation of the striatum, attenuating impairments of sensorimotor behaviour with a 36-38% increase in the maintenance of DARPP-32 / krox-24 expressing striatal neurons, reduced striatal atrophy and increased maintenance of striatonigral projections. Higher levels of BDNF however induced seizures and weight-loss highlighting the need to provide regulatable control over biotherapeutic protein expression. Continuous high-expression of BDNF or GDNF resulted in a downregulation of intracellular signal mediating proteins including DARPP-32, with AAV-GDNF not found to enhance the overall maintenance of striatal neurons. Neither of the anti-apoptotic factors provided significant protection of transduced striatal neurons but tended towards ameliorating QA-induced behavioural deficits, displaying behaviour – pathology correlations with the survival of parvalbumin-expressing neurons in the globus pallidus. The results of this thesis suggest BDNF as a promising putative biotherapeutic for HD, but emphasises the requirement to control expression following gene delivery, and for further elucidation of the physiological impact that enhanced expression of endogenous factors has on the host cells. Additionally the maintenance of neural networks beyond the caudate-putamen will be vital to ensuring efficient clinical outcomes for HD. / Auckland Medical Research Foundation. Foundation for Research, Science and Technology. The University of Auckland.

Progressive degeneration in the central nervous system (CNS) of Huntington’s disease (HD) patients is a relentless debilitating process, resulting from the inheritance of a single gene mutation. With limited knowledge of the underlying pathological molecular mechanisms, pharmaceutical intervention has to-date not provided any effective clinical treatment strategies to attenuate or compensate the neuronal cell death. Attention has therefore turned to biotherapeutic molecules and novel treatment approaches to promote restoration and protection of selectively vulnerable populations of neurons in the HD brain. Rapid advances in vectorology and gene-based medicine over the past decade have opened the way for safe and efficient delivery of biotherapeutics to the CNS. With numerous factors known to regulate the development, plasticity and maintenance of the mammalian nervous system many proteins have emerged as potential therapeutic agents to alleviate HD progression. This investigative study utilised gene delivery vectors derived from the non-pathogenic adeno-associated virus (AAV) to direct high-level expression of brain-derived neurotrophic factor (BDNF), glial cell-line derived neurotrophic factor (GDNF), Bcl-xL or X-linked inhibitor of apoptosis protein (XIAP) within the rodent striatum. Maintenance of the basal ganglia and functional behaviour deficits were assessed following excitotoxic insult of the striatum by quinolinic acid (QA), a neurotoxic model of HD pathology. Enhanced striatal expression of BDNF prior to QA-induced lesioning provided maintenance of the striosome-matrix organisation of the striatum, attenuating impairments of sensorimotor behaviour with a 36-38% increase in the maintenance of DARPP-32 / krox-24 expressing striatal neurons, reduced striatal atrophy and increased maintenance of striatonigral projections. Higher levels of BDNF however induced seizures and weight-loss highlighting the need to provide regulatable control over biotherapeutic protein expression. Continuous high-expression of BDNF or GDNF resulted in a downregulation of intracellular signal mediating proteins including DARPP-32, with AAV-GDNF not found to enhance the overall maintenance of striatal neurons. Neither of the anti-apoptotic factors provided significant protection of transduced striatal neurons but tended towards ameliorating QA-induced behavioural deficits, displaying behaviour – pathology correlations with the survival of parvalbumin-expressing neurons in the globus pallidus. The results of this thesis suggest BDNF as a promising putative biotherapeutic for HD, but emphasises the requirement to control expression following gene delivery, and for further elucidation of the physiological impact that enhanced expression of endogenous factors has on the host cells. Additionally the maintenance of neural networks beyond the caudate-putamen will be vital to ensuring efficient clinical outcomes for HD. / Auckland Medical Research Foundation. Foundation for Research, Science and Technology. The University of Auckland.

[Truncated abstract] There is limited intrinsic potential for repair in the adult human central nervous system (CNS). Dysfunction resulting from CNS injury is persistent and requires prolonged medical treatment and rehabilitation. The retina and optic nerve are CNSderived, and adult retinal ganglion cells (RGCs) and their axons are often used as a model in which to study the mechanisms associated with injury, neuroprotection and regeneration. In this study I investigated the effects of a variety of strategies on promoting RGC survival and axonal regeneration after optic nerve injury, including the use of reconstructed chimeric peripheral nerve (PN) grafts, gene therapy, and intraocular application of pharmacological agents and other factors . . . C3 transferase is an enzyme derived from Clostridium botulinum that inactivates Rho GTPase. Because SC myelin contains MAG and PN also contains CSPGs, I tested the effects of intraocular injection of a modified form of C3 (C3-11), provided by Dr Lisa McKerracher (CONFIDENTIAL data, under IP agreement with Bioaxone Therapeutic, Montreal) on RGC axonal regeneration into PN autografts. My results showed that there was significantly more RGC survival and axonal regeneration in PN autografts after repeated intraocular injection of C3. I also tested whether intraocular injections of CPT-cAMP and/or CNTF can act in concert with the C3 to further increase RGC survival and/or regeneration. Results showed that the effect of C3 and CPT-cAMP plus CNTF were synergistic and partially additive. The use of combination therapies therefore offers the best hope for robust and substantial regeneration. The overall results from my PhD project will help determine how best to reconstruct nerve pathways and use pharmacological interventions in the clinical treatment of CNS injury, hopefully leading to improved functional outcomes after neurotrauma.

Submitted by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2014-12-17T14:07:27Z No. of bitstreams: 2 adriano_prado_simao.pdf: 14603272 bytes, checksum: 6a0e234b9d484325125c4f251d688d0b (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2014-12-17T14:08:37Z (GMT) No. of bitstreams: 2 adriano_prado_simao.pdf: 14603272 bytes, checksum: 6a0e234b9d484325125c4f251d688d0b (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2014-12-17T14:08:31Z (GMT) No. of bitstreams: 2 adriano_prado_simao.pdf: 14603272 bytes, checksum: 6a0e234b9d484325125c4f251d688d0b (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2014-12-17T14:09:21Z (GMT) No. of bitstreams: 2 adriano_prado_simao.pdf: 14603272 bytes, checksum: 6a0e234b9d484325125c4f251d688d0b (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) / Made available in DSpace on 2014-12-17T14:09:21Z (GMT). No. of bitstreams: 2 adriano_prado_simao.pdf: 14603272 bytes, checksum: 6a0e234b9d484325125c4f251d688d0b (MD5) license_rdf: 22974 bytes, checksum: 99c771d9f0b9c46790009b9874d49253 (MD5) Previous issue date: 2013 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior (Capes) / Funda??o de Amparo ? Pesquisa do estado de Minas Gerais (FAPEMIG) / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico (CNPq) / Introdu??o: Recentemente, a vibra??o de todo o corpo (VTC) tem sido um m?todo de exerc?cio f?sico indicado para aumentar o desempenho e a capacidade f?sico-funcional de idosos com osteoartrite (OA) de joelho. No entanto, os mecanismos relacionados aos efeitos produzidos por essa modalidade ainda n?o foram completamente investigados. Objetivos: O objetivo deste estudo foi investigar os efeitos da adi??o de VTC aos exerc?cios de agachamento na concentra??o plasm?tica de marcadores inflamat?rios e no desempenho e capacidade funcionais de idosos com OA de joelho na fase remissiva da doen?a (Estudo 1) e investigar os efeitos da adi??o do treinamento de VTC ao exerc?cio de agachamento em mulheres idosas com OA de joelho na fase remissiva da doen?a nos seguintes par?metros: 1) for?a isom?trica do m?sculo quadr?ceps; 2) concentra??o plasm?tica de BDNF; e 3) na concentra??o salivar da raz?o testosterona/cortisol (Estudo 2). Investigar a rela??o entre os n?veis plasm?ticos e no l?quido sinovial do TNF-? e seus receptores sol?veis (sTNFR1 e sTNFR2) assim como de BDNF em idosos com OA de joelho e ainda verificar a rela??o destes com a gravidade da OA e o autorrelato de dor, rigidez e fun??o f?sica com o WOMAC (Western Ontario and McMaster University Osteoarthritis Index) em idosos com OA de joelho na fase inflamat?ria aguda (Estudo 3). Metodologia: No estudo 1, trinta e dois idosos com OA de joelho foram divididos em tr?s grupos: grupo que realizou exerc?cio de agachamento associado a plataforma vibrat?ria (grupo plataforma N=11), grupo que realizou exerc?cio de agachamento sem vibra??o (grupo agachamento N=10) e o grupo controle (N=11). Um programa estruturado de exerc?cios de agachamento foi executado tr?s vezes por semana em dias alternados por doze semanas nos grupos plataforma e agachamento. A concentra??o plasm?tica de receptores sol?veis de TNF-? (sTNFR1 e sTNFR2) foi analisada usando a t?cnica de ELISA. O WOMAC foi usado para avaliar o autorrelato da fun??o f?sica, dor e rigidez. O teste de caminhada de 6 minutos, a escala de Berg e o teste de velocidade da marcha foram utilizados para avaliar a fun??o f?sica. No estudo 2, foram selecionadas quinze mulheres idosas com idade maior ou igual a 60 anos que tinham sido diagnosticadas com OA em pelo menos um joelho. A interven??o consistiu de doze semanas seguidas de exerc?cios de agachamento, 3 vezes por semana. O protocolo de exerc?cio foi similar em ambos os grupos diferindo apenas da presen?a de vibra??o. J? no estudo 3 participaram vinte e sete idosos diagnosticados com osteoartrite de joelho e dezenove idosos saud?veis. Radiografias ?ntero-posteriores do joelho foram realizadas para determinar a gravidade da doen?a no joelho afetado. A classifica??o radiogr?fica da OA do joelho foi realizada utilizando os crit?rios Kellgren-Lawrence. Os n?veis de TNF-?, sTNFR1, sTNFR2 e BDNF no plasma e no l?quido sinovial foram determinados por ELISA. Resultados: No estudo 1, o grupo que realizou exerc?cios de agachamento na plataforma vibrat?ria mostrou diminui??o nas concentra??es plasm?ticas dos marcadores inflamat?rios sTNFR1 e sTNFR2 (p<0,001 e p<0,05, respectivamente), no autorrelato da dor (p<0,05), melhora no equil?brio (p<0,05) e na velocidade e dist?ncia caminhada (p<0,05 e p<0,001, respectivamente) comparado com o grupo controle. O teste de velocidade da marcha tamb?m apresentou aumento no grupo plataforma quando comparado ao grupo agachamento (p<0,01). Os resultados do estudo 2 demonstraram uma varia??o (?) positiva dos valores da for?a isom?trica muscular do quadr?ceps (p=0,02) e da concentra??o plasm?tica de BDNF (p=0,03) no grupo vibra??o ap?s o per?odo de interven??o. A varia??o (?) na raz?o testosterona/cortisol (T/C) n?o diferiu significativamente, entre os grupos (p=0,61). No estudo 3, os n?veis de BDNF no l?quido sinovial correlacionou-se significativamente com dor autorrelatada [WOMAC] (rs = 0,39, p=0,04). Com rela??o aos receptores sol?veis para TNF-?, observou-se uma diferen?a entre os n?veis de sTNFR1 e de sTNFR2, tanto no plasma quanto no l?quido sinovial em pacientes com OA do joelho (1091 ? 99,48 pg / mL versus 2249 ? 126,3 pg / mL e 2587 ? 66,12 pg / mL versus 2021 ? 107,0 pg / mL, respectivamente). Al?m disso, os n?veis de sTNFR1 no l?quido sinovial foram, negativamente, correlacionadas com a dor e a fun??o f?sica autorrelatada (rs -0,6785, p<0,0001 e rs -0,4194; p=0,03, respectivamente). Ao passo que, os n?veis de sTNFR2 no l?quido sinovial foram negativamente correlacionadas com dor e rigidez articular (rs -0,5433, p=0,01 e rs -0,4249; p=0,02, respectivamente). Conclus?es: Os resultados dos estudos supracitados indicam que a adi??o da vibra??o de todo o corpo ao treinamento com exerc?cio de agachamento, nas condi??es experimentais propostas, melhora o equil?brio est?tico e din?mico e o desempenho da marcha e ao mesmo tempo reduz a autopercep??o de dor e a concentra??o de marcadores inflamat?rios (sTNFR1 e sTNFR2) em idosos com OA de joelho na fase de remiss?o da doen?a. Al?m disso, a associa??o da vibra??o de todo o corpo ao exerc?cio de agachamento promove uma melhora na for?a muscular de membros inferiores em mulheres idosas com OA de joelho na fase de remiss?o da doen?a e proporciona uma resposta adaptativa na concentra??o de BDNF sem altera??o na rela??o muscular de anabolismo/catabolismo. J? os resultados da rela??o entre sist?mico e local, indicam que os n?veis de BDNF sist?micos est?o associados com o mecanismo da dor articular na OA de joelho. Com rela??o aos receptores sol?veis de TNF-?, evidenciou-se a presen?a de receptores sol?veis para TNF-? no l?quido sinovial de pacientes com OA prim?ria de joelho e a rela??o destes receptores com par?metros cl?nicos. / Tese (Doutorado) ? Programa Multic?ntrico de P?s-Gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2013. / ABSTRACT Introduction: Recently, whole body vibration (WBV) has been an alternative method of exercise that has been indicated to improve the physical performance of the elderly with osteoarthritis (OA) knee. However, the mechanisms related to the effects produced by this training mode have not been fully elucidated in the literature. Objectives: 1) To investigate the effects of the adittion of whole-body vibration to squat exercises on the plasma concentration of inflammatory markers and the functional performance of elderly individuals with knee OA remission phase (Study 1) and investigate the effects of WBV in addition to squat exercise training in elderly women with knee OA remission phase on the following parameters: 1) isometric strength of the quadriceps muscle; 2) BDNF plasma concentration; and 3) the testosterone/cortisol salivary concentration ratio (Study 2). 3). To analyze the concentrations of TNF-?, soluble receptors (sTNFR1 and sTNFR2) and BDNF in both plasma and synovial fluid of patients with inflammatory acute phase primary knee osteoarthritis during inflammatory acute phase, and to determine the possible correlations of plasma and synovial fluid TNF-?, soluble receptors (sTNFR1 and sTNFR2) and BDNF with the radiographic grading of knee OA and with self-reported pain, stiffness and physical function (Study 3). Methods: In study 1 thirty-two elderly subjects with knee osteoarthritis were divided into three groups [i.e., squat exercises on a vibratory platform (platform group N= 11), squat exercises without vibration (squat group N= 10) and the control group (N=11)]. The structured program of squat exercises in the platform and squat groups was conducted three times per week, on alternate days, for twelve weeks. The plasma concentration of TNF-? soluble receptors (sTNFR1 and sTNFR2) were analyzed using ELISA. The Western Ontario and McMaster University Osteoarthritis Index (WOMAC) questionnaire were used to evaluate self-reported physical function, pain and stiffness. The 6-minute walk test, the Berg balance scale, and gait speed were used to evaluate physical function. In study 2 the eligible patients were fifteen (15) elderly women ? 60 years of age who had been diagnosed with OA in at least one knee. The intervention consisted of uninterrupted squatting exercises for twelve weeks, 3x/week. The exercise protocol was similar in both groups differing only in the presence of vibration. In study 3 samples of plasma taken from the peripheral blood and of synovial fluid taken from the knee of patients with osteoarthritis (OA) were collected (n=27). Anteroposterior knee radiographs were taken to determine disease severity in the affected knee. Radiographic grading of OA in the knee was performed using the Kellgren-Lawrence criteria. Furthermore, plasma was collected from the peripheral blood of 19 healthy individuals, with no radiographic change in the hips and knees. The Western Ontario and McMaster University Osteoarthritis Index (WOMAC) questionnaire was used to evaluate self-reported physical function, pain and stiffness. ELISA measured the TNF-?, sTNFR1, sTNFR2 and BDNF levels in the plasma and synovial fluid. Results: In the study 1 the group that performed squat exercises on a vibratory platform, there were significant differences in plasma concentrations of the inflammatory markers sTNFR1 and sTNFR2 (p<0.001 and p<0.05, respectively), self-reported pain (p<0.05), balance (p<0.05) and speed and distance walked (p<0.05 and p<0.001, respectively) compared with the control group. The gait speed test also showed significant differences between the squat and platform groups (p<0.01). In the results of the study 2, the VG group demonstrated a significantly greater change (?) in IQMS values (p = 0.02) and in BDNF plasma concentrations (p = 0.03) after the intervention period compared with the EG group. The change (?) in T/C ratio showed no difference between the groups (p = 0.61). In the results of the study 3, the plasma BDNF levels significantly correlated with self-reported pain [WOMAC] (rs=0.39, p=0.04). According to soluble receptors to TNF-?, there was a difference between sTNFR1 and sTNFR2 levels in plasma as well as in synovial fluid in patients with knee (1091 ? 99,48 pg / mL versus 2249 ? 126,3 pg / mL e 2587 ? 66,12 pg / mL versus 2021 ? 107,0 pg / mL, respectively). Synovial fluid sTNFR1 levels were negatively correlated with pain and physical function self-reported (rs-0.6785, p<0.0001 and rs-0.4194, p=0.03, respectively). Synovial fluid sTNFR2 levels were negatively correlated with pain and joint stiffness (rs-0.5433, p=0.01 and rs-0.4249, p=0.02, respectively). Conclusions: The results of the above studies indicate that the addition of vibration training the whole body to squat exercise in the experimental conditions resulted in improvement in static and dynamic balance and gait performance and reduced the self-perception of pain and the concentration of inflammatory markers (sTNFR1 and sTNFR2) in elderly patients with knee OA. We also demonstrate that the combination of vibration training the whole body to squat exercise can promote an improvement in lower limb muscle strength in elderly women with knee OA and still provide an adaptive response to the concentration of BDNF compared with no change in muscle anabolism/catabolism. The results of the relationship between systemic and local concentration indicate that the systemic BDNF levels are associated with the mechanism of joint pain in knee OA. With respect to TNF-? soluble receptors, the findings demonstrated the presence of soluble receptors for TNF-alpha, particularly sTNFR1, in the synovial fluid of patients with primary knee OA and the relationship between these receptors and clinical parameters.

Submitted by Jos? Henrique Henrique (jose.neves@ufvjm.edu.br) on 2017-03-02T20:23:00Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) ana_cristina_resende_camargos.pdf: 3572613 bytes, checksum: ed282931c95236a0305016db8c75f026 (MD5) / Approved for entry into archive by Rodrigo Martins Cruz (rodrigo.cruz@ufvjm.edu.br) on 2017-03-06T11:37:07Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) ana_cristina_resende_camargos.pdf: 3572613 bytes, checksum: ed282931c95236a0305016db8c75f026 (MD5) / Made available in DSpace on 2017-03-06T11:37:08Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) ana_cristina_resende_camargos.pdf: 3572613 bytes, checksum: ed282931c95236a0305016db8c75f026 (MD5) Previous issue date: 2016 / A obesidade infantil ? considerada um dos maiores problemas de sa?de p?blica do mundo. Estudos demonstram altera??o do padr?o de secre??o de adipocinas, cortisol e do fator neurotr?fico derivado do c?rebro (BDNF), bem como inflama??o cl?nica cr?nica sublimiar e desequil?brio redox em crian?as com sobrepeso e obesidade na idade escolar. Al?m disso, o excesso de peso tamb?m est? associado a reduzido desenvolvimento cognitivo e motor. Apesar da literatura indicar que os primeiros 24 meses de vida representam um per?odo importante para o desenvolvimento da obesidade infantil, n?o existem evid?ncias se crian?as com sobrepeso e obesidade nesta faixa et?ria j? apresentam altera??es em par?metros neuroend?crino-inflamat?rios, com poss?vel impacto no desenvolvimento infantil. Dessa forma, os objetivos deste estudo foram: 1) analisar as concentra??es plasm?ticas de adipocinas e BDNF, as concentra??es s?ricas de cortisol e o estado redox de crian?as com sobrepeso/obesidade entre seis e 24 meses de idade; 2) avaliar o desenvolvimento cognitivo e motor de crian?as com sobrepeso/obesidade entre seis e 24 meses de idade e; 3) verificar associa??es entre os biomarcadores avaliados com o desenvolvimento cognitivo e motor nessa faixa et?ria. Foi realizado um estudo transversal com crian?as com sobrepeso/obesidade e eutr?ficas entre seis e 24 meses de idade, cadastradas nas Estrat?gias de Sa?de da Fam?lia. As concentra??es plasm?ticas de leptina, adiponectina, resistina, receptores sol?veis do fator de necrose tumoral 1 e 2 (sTNFR1 e sTNFR1), BDNF e as concentra??es s?ricas de cortisol foram mensuradas pelo m?todo ELISA. As quimiocinas foram mensuradas pela t?cnica cytometric bead arrays e o estado redox foi determinado por meio da detec??o das concentra??es de subst?ncias reativas ao ?cido tiobarbit?rico (TBARS), da atividade das enzimas catalase (CAT) e super?xido dismutase (SOD) e da habilidade de redu??o do ferro (FRAP). O desenvolvimento infantil foi avaliado por meio do instrumento Bayley Scales of Infant and Toddler Development, 3? edi??o (Bayley-III). Foi utilizado teste t para amostras independentes para comparar os grupos e correla??o de Pearson ou Spearman para verificar a associa??o entre as vari?veis. Al?m disso, foi utilizado um modelo de regress?o linear m?ltipla stepwise para verificar a associa??o entre os biomarcardores selecionados com o desenvolvimento cognitivo e motor. As concentra??es plasm?ticas de leptina (p=0,0001), adiponectina (p=0,0007), BDNF (p=0,003) e as concentra??es s?ricas de cortisol (p=0,048) foram significativamente superiores no grupo de crian?as com sobrepeso/obesidade. Em contrapartida, as crian?as deste grupo apresentaram menores concentra??es de TBARS (p=0,004) e menor atividade das enzimas antioxidantes CAT (p=0,045) e SOD (p=0,02). N?o foram encontradas diferen?as significativas nas concentra??es de quimiocinas, sTNFR1, sTNFR2, resistina e FRAP entre os grupos (p>0,05). Al?m disso, as crian?as do grupo sobrepeso/obesidade apresentaram menores escores de desenvolvimento cognitivo (p=0,03) e motor (p=0,04). Foi ainda encontrada associa??o significativa entre as concentra??es plasm?ticas de leptina e sTNFR1 com o escore composto cognitivo (p=0,001) e das concentra??es plasm?ticas de sTNFR1 com o escore composto motor (p=0,003). Todos esses resultados apontaram a presen?a de altera??es neuroend?crino-inflamat?rias em crian?as com sobrepeso/obesidade entre seis e 24 meses de idade. Al?m disso, embora a maior parte das crian?as com sobrepeso/obesidade apresentem desenvolvimento infantil dentro dos limites de normalidade, evidencia-se pior desempenho cognitivo e motor. Por fim, foi demonstrado que maiores concentra??es de sTNFR1 e menores concentra??es de leptina foram associadas com melhores desfechos de desenvolvimento infantil nessa faixa et?ria. / Tese (Doutorado) ? Programa Multic?ntrico de P?s-Gradua??o em Ci?ncias Fisiol?gicas, Universidade Federal dos Vales do Jequitinhonha e Mucuri, 2016. / Childhood obesity is one of the world?s most serious public health issues. Studies have demonstrated dysregulated secretion pattern of adipokines, cortisol, brain-derived neurotrophic factor (BDNF), as well as chronic low-grade inflammation and redox imbalance in school-age overweight or obese children. Overweight/obese infants also had lower cognitive and motor development scores. Although the literature points out that the first 24 months of life represent an important period for the development of childhood obesity, it is not known whether overweight/obese infants in this age interval present alterations in neuroendocrine inflammatory parameters, with possible impact on child development. Then, the objectives of this study were: 1) to analyze the plasmatic levels of adipokines and BDNF, serum cortisol and redox status in overweight/obese infants between 6 and 24 months of age; 2) to evaluate the cognitive and motor development in overweight/obese infants between 6 and 24 months of age and; 3) to verify the association of the biomarkers evaluated with cognitive and motor development in this age interval. A cross-sectional study was conducted with infants with overweight/obesity and normal-weight between 6 and 24 months enrolled in Family Health Strategies. Plasma leptin, adiponectin, resistin, BDNF, soluble tumour necrosis factor receptors 1 and 2 (sTNFR1, sTNFR2), and serum cortisol levels were measured using conventional ELISA kits. Plasma chemokines were measured using the cytometric bead arrays kit, and oxidative stress was assessed by thiobarbituric acid reactive substances (TBARS) concentration, enzymes catalase (CAT) and dismutase superoxide (SOD) activity, as well by ferric reducing antioxidant power (FRAP). Infant development was performed using the Bayley Scales of Infant and Toddler Development, 3rd edition (Bayley-III). A t test for independent samples was performed to compare the groups and Pearson and Spearman correlation was used to verify the association between parameters. Multiple linear stepwise regression models were utilized to verify the association between the biomarkers selected and cognitive and motor composite scores. Plasma levels of leptin (p=0.0001), adiponectin (p=0.0007), BDNF (p=0.003) and serum cortisol (p=0.048) were significantly higher in overweight/obese infants. In contrast, concentration of TBARS (p=0.004), CAT (p=0.045) and SOD activity (p=0.02) were lower in overweight/obese infants. There were no differences in the levels of chemokines, sTNFR1, sTNFR2, resistin and FRAP between groups (p>0.05). Moreover, overweight/obese infants had lower cognitive (p=0.03) and motor (p=0.04) development scores than normal-weight infants. A significant association of plasma leptin and sTNFR1 levels with cognitive composite scores (p=0.001) were found and plasma sTNFR1 levels were associated with motor composite scores (p=0.003). All these results point out neuroendocrine inflammatory response changes in overweight/obese infants between 6 and 24 months. Moreover, although most of overweight/obese infants have presented cognitive and motor development within normal limits, there is evidence of worse cognitive and motor performance. Finally, high sTNFR1 and low leptin levels were associated with increase developmental outcomes in infants in this age interval.

Submitted by VÍTOR BONETTI VALENTE null (vitorbvalente89@hotmail.com) on 2016-10-14T20:59:55Z No. of bitstreams: 1 Dissertação - Vitor B. Valente_Versão Definitiva.pdf: 1948563 bytes, checksum: 03b2d1c12fa8b8b2c7b0c39faf9bb490 (MD5) / Approved for entry into archive by Juliano Benedito Ferreira (julianoferreira@reitoria.unesp.br) on 2016-10-19T19:14:01Z (GMT) No. of bitstreams: 1 valente_vb_me_arafo_par.pdf: 678709 bytes, checksum: 264cdeca2ea5cfd220b36a541cd5ae2c (MD5) / Made available in DSpace on 2016-10-19T19:14:01Z (GMT). No. of bitstreams: 1 valente_vb_me_arafo_par.pdf: 678709 bytes, checksum: 264cdeca2ea5cfd220b36a541cd5ae2c (MD5) Previous issue date: 2016-09-02 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Evidências mostram que os hormônios relacionados ao estresse podem influenciar a progressão do câncer, mas o papel destes mediadores sobre o processo de carcinogênese no microambiente tecidual, em condições naturais, é pouco compreendido. Neste estudo, nós utilizamos um modelo de carcinogênese bucal em ratos para testar a hipótese de que os níveis de hormônios relacionados ao estresse no microambiente tecidual em condições naturais (sem estresse) pré-indução carcinogênica influenciam a ocorrência e progressão do carcinoma espinocelular (CEC) de língua. Quarenta e oito ratos machos Wistar foram submetidos a uma biópsia de tecido lingual normal previamente à indução carcinogênica e os níveis teciduais de norepinefrina, corticosterona, ACTH e BDNF foram mensurados. Três semanas depois os animais foram tratados com o carcinógeno químico 4-nitroquinolina-1-óxido (4NQO) por 20 semanas e a ocorrência de CEC ou Leucoplasia (lesão precursora do CEC) na língua foi analisada microscopicamente. Níveis basais aumentados pré-carcinogênese de norepinefrina e BDNF e níveis reduzidos de corticosterona foram preditivos para ocorrência de CEC. Níveis basais elevados de norepinefrina foram associados à uma expressão reduzida de RNAm para CDKN2a-p16 nos CECs. Níveis teciduais de corticosterona e BDNF nas leucoplasias e corticosterona no CEC foram significativamente mais elevados em relação a mucosa normal pré-carcinogênese. Níveis elevados de norepinefrina no microambiente dos CECs foram associados a um maior volume e espessura do tumor. Da mesma forma, níveis elevados de norepinefrina, ACTH e BDNF no CEC foram associados a uma menor intensidade do infiltrado linfoplasmocitário subjacente ao tumor. Além disso, maior expressão de RNAm para IL-6 no CEC foi correlacionada à níveis elevados de corticosterona pós-carcinogênese. Este estudo mostra as primeiras evidências in vivo de que os níveis basais de hormônios do estresse no microambiente do tecido normal podem ser preditivos para a incidência do câncer quimicamente induzido. Além disso, a ação do carcinógeno pode modular os níveis hormonais no microambiente tecidual e estes podem estar associados à progressão do tumor. Em suma, estes dados sugerem que a susceptibilidade ao início e progressão do carcinoma quimicamente induzido pode ser diretamente influenciada por diferenças individuais endócrinas no microambiente pré-carcinogênese.

Introdução: Em crianças e adolescentes, o Transtorno de Humor Bipolar (THB) é associado a danos devastadores no desenvolvimento. O Transtorno de Déficit de Atenção/Hiperatividade (TDAH), caracterizado por sintomas de desatenção, hiperatividade e impulsividade também causa prejuízo funcional significativo. O diagnóstico diferencial entre os dois transtornos é puramente clínico e, até o momento, há poucos estudos avaliando diferenças neurobiológicas. Diversas pesquisas sugerem a participação do Fator Neurotrófico Derivado do Cérebro (BDNF), cujo papel não foi elucidado em crianças e adolescentes com THB e com TDAH. Apesar das altas taxas de comorbidade entre o THB e o TDAH, de uma pior resposta ao tratamento e de um pior funcionamento psicossocial quando da comorbidade, apenas dois estudos avaliaram a resposta ao tratamento especificamente neste grupo de pacientes. Objetivos: O objetivo principal deste trabalho é avançar no entendimento do papel do BDNF na psicopatologia do THB e do TDAH, visando sua diferenciação. O tratamento da comorbidade com estimulantes tambem foi estudado. Métodos: A transmissão do alelo Val66 do BDNF foi avaliada em famílias de crianças e adolescentes com THB e TDAH, assim como o efeito do gene no nível sérico da proteína do BDNF entre os grupos. Foi realizada a comparação dos níveis séricos entre pacientes com THB em comorbidade com TDAH e TDAH isolado. Um estudo clínico cruzado com estimulantes foi realizado em crianças e adolescentes com THB e TDAH em comorbidade que tinham apresentado remissão dos sintomas de humor com o uso de aripiprazol, mas persistência dos sintomas de TDAH. Os sintomas de mania, depressão, desatenção e hiperatividade foram acompanhados ao longo de quatro semanas de tratamento, duas com placebo ou metilfenidato e duas com o tratamento inverso. Resultados: Na investigação da transmissão do gene do BDNF em crianças e adolescentes não foi detectada diferença significativa na transmissão do alelo Val66. Tampouco foi observada diferença significativa nos níveis séricos da proteína do BDNF entre os pacientes com THB em comorbidade com TDAH, quando comparados aos pacientes com TDAH e controles. No estudo cruzado com crianças e adolescentes com THB em comorbidade com TDAH, não houve diferenças entre os grupos com placebo ou estimulantes na resposta ao tratamento nos sintomas de TDAH. Os sintomas de humor mantiveram-se estáveis a despeito do uso de metilfenidato. Conclusões: Os achados quanto ao gene do BDNF não sugerem sua participação na neurobiologia do THB ou no TDAH, ou que devido à herança poligênica característica dos transtornos mentais, sua participação seja pequena. O achado de diferença significativa entre os níveis séricos do BDNF de crianças e adolescentes com THB+TDAH e TDAH indica que esse tema deve ser mais estudado, e, caso seja também encontrado de forma consistente por outros grupos, possa vir a ser utilizado como marcador biológico na diferenciação diagnóstica entre essas condições. No estudo de tratamento da comorbidade entre THB e TDAH, a ausência de resposta ao metilfenidato nos sintomas de TDAH em pacientes que apresentam a comorbidade reforça a evidência de que há pior resposta ao tratamento neste grupo, dado o elevado tamanho de efeito do metilfenidato no tratamento do TDAH em isolado O estudo de fatores biológicos para um melhor entendimento da psicopatologia e conseqüente diferenciação dos transtornos mentais tem extrema relevância porque a identificação destes fatores pode auxiliar na elaboração de tratamentos mais precisos, urgentemente necessários devido às graves conseqüências do THB e do TDAH nas vidas dos pacientes e de suas famílias. A criação de um programa específico para Crianças e Adolescentes com Transtorno Bipolar (ProCAB) e de uma linha de pesquisa com foco na etiologia e tratamento possibilitam uma constante geração de conhecimento nesta área, onde poucos estudos estão disponíveis. / Introduction: Bipolar Disorder (BD) in children and adolescents is associated to devastating developmental deficits. Attention-Deficit/Hyperactivity Disorder (ADHD), characterized by inattention, hyperactivity, and impulsivity, also promotes significant impairment. Differential diagnosis between both conditions is purely clinical – currently, there are scarce investigations on neurobiological differences. Several studies suggest the participation of the Brain-Derived Neurotrophic Factor (BDNF) in these disorders, whose role has not been elucidated in BD and ADHD in children and adolescents. Despite high comorbidity rates between BD and ADHD, worse psychosocial functioning, and worse response to treatment, only two studies addressed treatment response specifically in this group of patients. Objectives: promote advances in understanding the role of BDNF in the psychopathology of BD and ADHD. The treatment of the comorbidity was also studied. Methods: Transmission of the Val66 allele at the BDNF was assessed in children and adolescents with BD and ADHD, as well as the effect of the gene on the serum levels of BDNF protein in both conditions. BDNF serum levels were compared between patients with BD comorbid with ADHD, and ADHD. A crossover clinical trial with stimulants and placebo was performed with children and adolescents preseting BD and comorbid ADHD. Manic, depressive, inatention and hyperactivity symptoms were assessed along a 4-week treatment, 2 weeks in each treatment arm (placebo or stimulants). Results: There was no significant transmission of the Val66 allele at the BDNF gene in children and adolescents with BD or ADHD. A significant difference in BDNF protein serum levels between BD+ADHD when compared to ADHD alone and controls. In the crossover trial with children and adolescents with BD and comorbid ADHD, we did not observe differences between the placebo and stimulant treatment groups in the response of ADHD symptoms. Mood symptoms remained stable despite the use of methylphenidate. Conclusions: our results regarding the BDNF gene do not suggest its participation in the neurobiology of BD or ADHD, or that due to the polygenic characteristic of mental disorders, that this gene confers a only a small risk, undetectable in our sample. The finding of a significant difference in BDNF serum levels between BD comorbid with ADHD, and ADHD alone warrants further investigation, and in case replication studies with larger samples from other groups are positive, BDNF serum levels might be used as a biological marker in the diagnostic difference between these conditions. In the investigation of the treatment of the comorbidity between BD and ADHD, the absence of different responses between placebo and methylphenidate in ADHD symptoms strengthens the evidence that there is a worse response to treatment in this group, given the large effect size of methylphenidate response in the treatment of ADHD alone. The quest for biological markers for a better understanding of the psychopathology and subsequent differentiation of mental disorders is extremely relevant. The identification of these factors may facilitate the creation of more accurate treatment regimens, urgently needed due to the severe developmental consequences of BD and ADHD in the patients’ and families’ lives. In this sense, the creation of a specific outpatient program for children and adolescent BD (ProCAB), a research line with focus on risk factors and treatment, will enable a Constant generation of knowledge in this área, where scarce data is available.