No description available.

Thesis (Ph. D.)--Duke University, 2008.

Made available in DSpace on 2015-12-06T23:44:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2006 / Objetivos: Estudar em ratos albinos portadores de diabetes melito induzido por estreptozotocina: 1. Valores plasmáticos de homocisteína. 2. Concentrações plasmáticas de fator von Willlebrand. 3. Valores séricos de colesterol total e frações e triglicérides. 4. Possíveis correlações entre valores plasmáticos de homocisteína, fator von Willebrand, colesterol total e frações e triglicérides. Métodos: 35 ratos (rattus norvegicus albinus), machos, adultos (peso 180-200g), com glicemias aferidas foram randomizados em três grupos: 1. Controle (n=10) - não receberam droga ou veículo; 2. “Sham” (n=10) - receberam solução (tampão citrato 0.1M, pH4.5) veículo da estreptozotocina e 3. Diabetes (n=15) - receberam estreptozotocina (Sigma® ) para indução do diabetes melito (60 mg/kg de peso) em dose única; via intraperitoneal, diluída em 0.3mL de solução veículo. Foram considerados diabéticos todos os ratos cujos valores de glicemia foram iguais ou superiores a 250mg/dL. Após 08 semanas de indução do diabetes melito os ratos foram pesados, as glicemias aferidas e anestesiados com ionembutal (Sigma® ) via intraperitoneal (50mg/kg peso). Colheu-se sangue da artéria aorta abdominal para determinação dos valores de homocisteína plasmática total seguindo metodologia descrita, através de ensaio HPLC (Shimadzu® ). O sangue restante foi centrifugado e o soro e o plasma aliquotados e congelados até determinação do fator von Willebrand (técnica de ELISA) e concentrações séricas de glicose de jejum final, colesterol total e frações HDL-, LDL- e VLDL-colesterol, triglicérides e creatinina. Os resultados foram expressos em média + desvio padrão. Para análise estatística utilizou-se a análise de variância (ANOVA), seguido do teste de comparações múltiplas de Tukey e quando necessário o teste Brown Forsythe, seguido do procedimento de comparações múltiplas de Dunnett. Para a construção dos diagramas de dispersão das variáveis calculou-se os coeficientes de correlação de Pearson. Resultados: O modelo foi reprodutível em 100% dos animais. A média dos valores de glicemia inicial foi: 88,7±5,9mg/dL (controle); 88,9±8,2mg/dL (“sham”) e 85,1±5,2mg/dL (diabetes). Através da ANOVA não houve diferença das médias de glicemia inicial entre os grupos (p=0,24). A média da glicemia final foi: 85,0±7,1mg/dL (controle); 80,9±5,0mg/dL (”sham”) e 353,5±98,2mg/dL (diabetes). Houve diferença estatisticamente significante entre o grupo diabetes e os demais (p<0,01). A média das concentrações plasmáticas de homocisteína foi: 7,9±2,3µmol/L (controle); 8,6±2,2µmol/L (“sham”) e 6,1±1,3µmol/L (diabetes), com diferença entre os grupos (p<0,01). A média dos valores do fator von Willebrand foi 0,15±0,3U/L (controle), 0,16±0,2U/L (“sham”) e 0,18±0,4U/L (diabetes), com diferença entre os grupos (p<0,05). Os valores de colesterol total tiveram médias de: 123,9±40,8mg/dL (controle); 107,0±38,6mg/dL (“sham”) e 87,5±5,9mg/dL (diabetes). A ANOVA mostrou diferença entre os grupos (p<0,05). No grupo diabetes houve correlação inversa entre glicemia final e ganho de peso, homocisteína e colesterol total, homocisteína e fração VLDL-colesterol e homocisteína e triglicérides. Conclusões: Neste estudo, utilizando como modelo biológico o diabetes melito induzido por estreptozotocina em ratos albinos, nas condições de experimento apresentadas, torna-se lícito concluir: 1. Os ratos diabéticos apresentaram valores menores de homocisteína. 2. O grupo diabetes apresentou valores maiores de fator von Willebrand. 3. Houve correlação inversa entre homocisteína e colesterol total, homocisteína e fração VLDL-colesterol e homocisteína e triglicérides. 4. Não houve correlação entre glicemia final e homocisteína, glicemia final e fator von Willebrand e homocisteína e fator von Willebrand nos ratos diabéticos. 5. Houve correlação inversa entre ganho de peso e glicemia final nos ratos diabéticos. / Purpose: To determine plasma levels of homocysteine, von Willebrand factor, triglycerides and total cholesterol and fractions, in rats with diabetes induced by streptozotocin and evaluate possible correlations among these parameters. Methods: Adult male norvegicus albino rats (n=35), weigh (180-200g) were randomized into three groups: treated group (n=15), with diabetes induced by streptozotocin; sham group (n=10), treated with saline solution and normal control group (n=10), no treated. Initial fasting glucose was determinated before diabetes induction. Diabetes was induced by a single bolus intraperitoneal injection of streptozotocin, 60mg/kg/dose, diluted in citrate buffer (0.1M, pH4.5). Diabetes was confirmed by blood glucose levels ≥250mg/dL. Eight weeks after diabetes induction, animals were weighted and blood samples were collected from abdominal aorta for plasma levels of total homocystein, von Willebrand factor, final fasting glucose, total cholesterol and fractions (HDL-, LDL- and VLDL-cholesterol), triglycerides and creatinin. The results were expressed as the mean+SD. Data were analyzed using analysis of variance (ANOVA) followed by Tukey test or Brown Forsythe test followed by Dunnett test. Pearson test was used to correlate the parameters. The level of significance was set at p<0.05. Results: Disease model reproducibility was observed in 100% of tested animals. Mean plasma levels of initial fasting glucose according to animal group were: 85.1±5.2mg/dL for treated group, 88.9±8.2mg/dL for sham and 88.7±5.9mg/dL for control group. Statistical analysis (ANOVA) showed no significant difference among the three groups (p=0.24). Mean plasma levels of final fasting glucose, according to animal group were: 353.5±98.2mg/dL for treated group, 80.9±5.0mg/dL for sham and 85.0±7.1mg/dL for normal control. Statistical analysis showed that the difference between the treated group and the two other groups was statistically significant (p<0.01). Mean plasma concentration of homocystein according to animal groups was: 6.1±1.3µmol/L for treated group, 8.6±2.2µmol/L for sham and 7.9±2.3µmol/L for control group. Statistical analysis showed a significant difference in the three groups (p<0.01). Mean values for von Willebrand factor were: 0.18±0.4U/L for treated group, 0.16±0.2U/L for sham group and 0.15±0.3U/L for control group. Significant statistical difference was found among the three groups (p<0.05). Mean plasma levels of total cholesterol according to animal groups were: 87.5±5.9mg/dL for treated group, 107.0±38.6mg/dL for sham and 123.9±40.8mg/dL for control group. Significant statistical difference was found among the three groups (p<0.05). There was no statistical significant difference in the mean values of cholesterol fraction (LDL, HDL, VLDL), triglycerides and creatinin among the three groups. A negative correlation between final plasma glucose and weight gain, homocystein and total cholesterol, homocystein and VLDL-cholesterol and homocystein and triglycerides was found in the treated (diabetes) group. Conclusions: In this study, results from streptozotocin-induced diabetes rats showed: 1. Lower levels of homocystein; 2. Higher levels of von Willebrand factor; 3. Negative correlation between homocystein and total cholesterol; 4. No correlations between final fasting glucose and homocystein, final fasting glucose and von Willebrand factor and homocystein and von Willebrand factor; 5. Negative correlation between weight gain and final fasting glucose. / BV UNIFESP: Teses e dissertações

Introdução: A esquizofrenia é um transtorno mental grave, que com frequência leva a uma deterioração progressiva, que mais recentemente tem sido descrita como uma doença sistêmica, com alterações progressivas cerebrais e corporais, e em parte por causa disto, associada a menor expectativa de vida, em média 20 anos menos que a população geral não afetada. Apesar de não terem especificidade, já foram determinados diferentes marcadores inflamatórios e oxidativos envolvidos; em contrapartida, pouco tem sido descrito sobre marcadores sistêmicos da esquizofrenia. A experiência clínica mostra que os pacientes crônicos vão ficando com pele, musculatura, e postura diferente dos não doentes, porém não há registro de estudo buscando estudar a postura, apesar de sabermos que a mesma depende da interação de inúmeros sistemas, incluindo o estado inflamatório do sujeito. Objetivo: Definir padrões de postura dentro de parâmetros básicos (postura cifolordótica - PCL, postura relaxada ou desleixada - PRD, postura escoliótica - PE) em pacientes esquizofrênicos e correlacionar com fase de doença (inicial-tardia) e com marcadores de resposta inflamatória imediata (proteína C reativa - PCR) e tardia (fator de von Willenbrand - FvW). Método: Estudo transversal, por amostra de conveniência, aprovado pelo comitê de ética do Hospital de Clínicas de Porto Alegre (número 110083 HCPA). Foram recrutados 40 indivíduos com diagnóstico CID-10 e DSMIVTR de esquizofrenia, forma estabilizada, em tratamento ambulatorial no HCPA. Os pacientes foram subdivididos em 2 subgrupos, de acordo com o estágio da doença (estágio inicial n=15 menos de 10 anos do 1° surto, estágio tardio n=25 10 anos ou mais do 1° surto). O grupo controle (n=26) foi recrutado através de uma rede social (Facebook®). Todos os indivíduos foram submetidos à biofotogrametria pelo método SAPO® para avaliar a postura. O nível de significância adotado foi de 5% para todas as variáveis e as análises foram realizadas no programa Statistical Package for the Social Sciences (SPSS, versão 18.0). Resultados: Não houve perdas; em relação à postura o grupo estágio inicial apresentou 15 ângulos com diferenças significativas quando comparado aos valores de referência, enquanto no grupo estágio tardio apenas 7 ângulos foram significativos; já na comparação com o grupo controle, apenas 6 ângulos foram significativos num total de 19. Os marcadores inflamatórios (PCR e FvW) não foram significativos em comparação aos estágios inicial e tardio da doença, porém a PCR apresentou correlação com a gravidade da doença e o FvW com um ângulo postural da protusão da cabeça. A variável dor também apresentou correlação com 5 ângulos posturais, 2 da coluna e 3 dos membros inferiores; além disso o grupo estágio tardio teve maior prevalência de dor quando comparado ao estagio inicial. Conclusões: Existe um padrão postural comum na esquizofrenia caracterizado principalmente por protusão da cabeça, hiperlordose e escoliose, sendo que no início da doença é mais agravante e no estágio mais tardio se estabiliza. Isto pode ser explicado por dois fenômenos: aumento de peso e o enfraquecimento muscular. Adicionalmente, temos a influência da dor e dos fatores inflamatórios, onde a PCR se associou à gravidade da doença, mas não à postura; já o FvW e a dor se associaram aos ângulos posturais, demonstrando suas influências nessa doença. Isto sugere que existe uma maior atividade patológica no início da doença, não só no cérebro, mas também no organismo, incluindo músculos e tecido adiposo. A hiperlordose parece estar mais associada à obesidade e tecido adiposo, enquanto que a extensão da cabeça e escoliose mais a alterações de musculatura. Todos estes achados juntos, se confirmados em amostras maiores e mais heterogêneas, irão nos auxiliar em futuras condutas no tratamento destas alterações posturais características deste transtorno (hiperlordose lombar, anteriorização da cabeça, diminuição da cifose torácica), além de estimular busca de novos meios de intervenções com o enfoque nos diferentes estágios da doença. / Introduction: Schizophrenia is a severe mental disorder that leads to progressive deterioration that more recently has been described as a systemic disease with progressive changes in the brain and body, and partly because of this, associated with lower life expectancy, in average 20 years less than the not affected general population. Although not specific, it has already been determined different inflammatory and oxidative markers involved, however little has been described about systemic markers of schizophrenia. Clinical experience shows that chronic patients are presenting skin, muscles, and posture different from the non patients, but there are no records of studies seeking to study the posture, even though we know that it depends on the interaction of many systems, including the inflammatory state of the subject. Objective: To define patterns of posture within basic parameters (PCL, PRD, PE) in schizophrenic patients and correlate with illness stage and with inflammatory markers of immediate (CRP) and delayed (vWF) response. Method: Cross-sectional study of a convenience sample approved by the ethics committee of the Hospital de Clinicas de Porto Alegre (HCPA). Forty patients with stabilized schizophrenia were recruited in the clinic treatment of HCPA, and analyzed according to 2 subgroups of the stage of the illness (early stage n = 15 less than 10 years from the 1st outbreak, late stage n = 25 10 years or more from the 1st outbreak). The control group (n = 26) were recruited through a social network (Facebook ®). All subjects underwent photogrammetry by SAPO ® method to evaluate posture. The level of significance was set at 5% for all variables and the analyzes were performed using the Statistical Package for the Social Sciences (SPSS, version 18.0). Results: there was no loss, in relation to posture the initial stage group showed 15 angles with significant differences when compared with the reference values, while the late stage group only 7 angles were significant, as compared with the control group only 6angles were significant in a total of 19, the inflammatory markers (CRP and vWF) were not significant in comparison to the initial and late stages of the disease, but the CRP showed correlation with disease severity and the vWF with a postural angle of the protrusion of the head. The pain variable also showed correlation with 5 postural angles, 2 of the column and 3 of the lower limbs, besides the late stage group had a higher prevalence of pain when compared to the early stage. Conclusions: There is a common postural pattern in schizophrenia characterized primarily by protrusion of the head, hyperlordosis and scoliosis, and that early in the illness is more aggravating and in the later stage it stabilizes. And it can be explained by two phenomena: weight gain and muscle weakness. Additionally, we have the influence of pain and inflammatory factors, where the CRP was associated with disease severity but not the posture, but the vWF and the pain were associated with postural angles demonstrating their influences in this disease. This suggests that there is increased pathological activity in the onset of the disease, not only in the brain but also in the body including muscles and adipose tissue. The hyperlordosis appears to be most strongly associated with obesity and adipose tissue, whereas the length of the head and scoliosis more to changes in muscles. All together these findings, if confirmed in larger and more heterogeneous samples will help us in future conduct in treating these postural changes characteristic of this disorder (lumbar hyperlordosis, forward head posture, decreased thoracic kyphosis), besides stimulating the search for new means of interventions with the focus on different stages of the disease.

Made available in DSpace on 2014-06-11T19:33:26Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-12-10Bitstream added on 2014-06-13T20:05:17Z : No. of bitstreams: 1 mattoso_crs_dr_botfmvz.pdf: 656719 bytes, checksum: d01581a0d1caa64d2fc80ce00231b862 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / A Doença de von Willebrand (DvW) é o distúrbio hemostático hereditário mais comum nos homens e também nos cães. A DvW já foi diagnosticada em mais de 54 raças de cães nos EUA. A DvW é causada por um defeito quantitativo e/ou qualitativo do Fator de von Willebrand (FvW), e nos cães é dividida em 3 tipos, tipo I, II e III. O diagnóstico da DvW está baseado na quantificação do FvW plasmático e testes de função plaquetária dependentes do FvW. Esse estudo foi dividido em duas partes, avaliação da prevalência da DvW (parte I) e avaliação das alterações do FvW em cadelas gestantes, e durante diferentes estágios do ciclo reprodutivo, em cadelas não gestantes (parte II). Os objetivos da parte I foram determinar a prevalência da Doença de von Willebrand (DvW) em cães da região de Botucatu, São Paulo, Brasil, e avaliar os testes laboratoriais para diagnóstico desta doença. Foram utilizados 350 cães de variadas idades e raças, de ambos os sexos. Os animais utilizados no estudo não tinham histórico ou evidências clínicas de alterações hemostáticas. Foram realizados os seguintes testes: antígeno do Fator de von Willebrand (vWF:Ag), Tempo de sangramento da Mucosa Oral (TSMO), Tempo de tromboplastina parcial ativada (TTPa) e atividade do Fator VIII (FVIII). A prevalência encontrada para Doença de von Willebrand em cães da região de Botucatu foi de 1,43%. A determinação do vWF:Ag foi o melhor teste laboratorial para diagnosticar a DvW. Os objetivos da parte II foram a avaliação das alterações encontradas na concentração do FvW em cadelas gestantes, e também durante diferentes estágios do ciclo reprodutivo, observados após a gestação, em animais portadores e não portadores para a DvW, além da avaliação de correlação entre FvW e cortisol. Foram utilizadas 7 cadelas portadoras (GI) e 9 não portadoras (GII) para DvW. Os animais foram... / Von Willebrand disease (vWD) is the most common inherited hemostatic disturbance affecting men and also dogs. It has already been diagnosed in more than 54 dog breeds in the USA. vWD is caused by a quantitative and/or qualitative defect in von Willebrand Factor (vWF); in dogs, it is divided into types I, II and III. The diagnosis of vWD is based on plasma vWF quantification and vWF-dependent platelet function tests. This study was divided in two sections, evaluation of vWF prevalence (Section I) and evaluation of vWF concentrations in pregnant bitches, and during various reproductive cycle stages in nonpregnant bitches (section II). The aims of (section I) were to assess the prevalence of von Willebrand disease in dogs from the region of Botucatu, São Paulo State, Brazil, and to evaluate diagnostic tests. The study included 350 dogs of various ages, different breeds, and both sexes. Dogs included in the study had no historical or clinical evidence of abnormal bleeding. von Willebrand factor antigen (vWF:Ag), Buccal mucosal bleeding time (BMBT), Activated partial thromboplastin time (aPTT), and Factor VIII activity (FVIII) were evaluated in their ability to diagnose vWD. The prevalence of vWD in dogs was 1.43% in the Botucatu region of Brazil. Determination of vWF:Ag was the best laboratory test to diagnose vWD. The aims of section II were the evaluation of vWF concentrations in pregnant bitches, and during various reproductive cycle stages in non-pregnant bitches, observed after the pregnancy in vWD affected and unaffected bitches, over there of the correlation among vWF and cortisol. Were used seven bitches affected (GI) and nine unaffected (GII) to vWD. Animals were evaluated during the pregnancy, parturition, lactation, and during various reproductive cycle stages in non-pregnant bitches. vWF concentration did not change during reproductive cycle stages in non-pregnant bitches... (Complete abstract click electronic access below)

Introdução: A esquizofrenia é um transtorno mental grave, que com frequência leva a uma deterioração progressiva, que mais recentemente tem sido descrita como uma doença sistêmica, com alterações progressivas cerebrais e corporais, e em parte por causa disto, associada a menor expectativa de vida, em média 20 anos menos que a população geral não afetada. Apesar de não terem especificidade, já foram determinados diferentes marcadores inflamatórios e oxidativos envolvidos; em contrapartida, pouco tem sido descrito sobre marcadores sistêmicos da esquizofrenia. A experiência clínica mostra que os pacientes crônicos vão ficando com pele, musculatura, e postura diferente dos não doentes, porém não há registro de estudo buscando estudar a postura, apesar de sabermos que a mesma depende da interação de inúmeros sistemas, incluindo o estado inflamatório do sujeito. Objetivo: Definir padrões de postura dentro de parâmetros básicos (postura cifolordótica - PCL, postura relaxada ou desleixada - PRD, postura escoliótica - PE) em pacientes esquizofrênicos e correlacionar com fase de doença (inicial-tardia) e com marcadores de resposta inflamatória imediata (proteína C reativa - PCR) e tardia (fator de von Willenbrand - FvW). Método: Estudo transversal, por amostra de conveniência, aprovado pelo comitê de ética do Hospital de Clínicas de Porto Alegre (número 110083 HCPA). Foram recrutados 40 indivíduos com diagnóstico CID-10 e DSMIVTR de esquizofrenia, forma estabilizada, em tratamento ambulatorial no HCPA. Os pacientes foram subdivididos em 2 subgrupos, de acordo com o estágio da doença (estágio inicial n=15 menos de 10 anos do 1° surto, estágio tardio n=25 10 anos ou mais do 1° surto). O grupo controle (n=26) foi recrutado através de uma rede social (Facebook®). Todos os indivíduos foram submetidos à biofotogrametria pelo método SAPO® para avaliar a postura. O nível de significância adotado foi de 5% para todas as variáveis e as análises foram realizadas no programa Statistical Package for the Social Sciences (SPSS, versão 18.0). Resultados: Não houve perdas; em relação à postura o grupo estágio inicial apresentou 15 ângulos com diferenças significativas quando comparado aos valores de referência, enquanto no grupo estágio tardio apenas 7 ângulos foram significativos; já na comparação com o grupo controle, apenas 6 ângulos foram significativos num total de 19. Os marcadores inflamatórios (PCR e FvW) não foram significativos em comparação aos estágios inicial e tardio da doença, porém a PCR apresentou correlação com a gravidade da doença e o FvW com um ângulo postural da protusão da cabeça. A variável dor também apresentou correlação com 5 ângulos posturais, 2 da coluna e 3 dos membros inferiores; além disso o grupo estágio tardio teve maior prevalência de dor quando comparado ao estagio inicial. Conclusões: Existe um padrão postural comum na esquizofrenia caracterizado principalmente por protusão da cabeça, hiperlordose e escoliose, sendo que no início da doença é mais agravante e no estágio mais tardio se estabiliza. Isto pode ser explicado por dois fenômenos: aumento de peso e o enfraquecimento muscular. Adicionalmente, temos a influência da dor e dos fatores inflamatórios, onde a PCR se associou à gravidade da doença, mas não à postura; já o FvW e a dor se associaram aos ângulos posturais, demonstrando suas influências nessa doença. Isto sugere que existe uma maior atividade patológica no início da doença, não só no cérebro, mas também no organismo, incluindo músculos e tecido adiposo. A hiperlordose parece estar mais associada à obesidade e tecido adiposo, enquanto que a extensão da cabeça e escoliose mais a alterações de musculatura. Todos estes achados juntos, se confirmados em amostras maiores e mais heterogêneas, irão nos auxiliar em futuras condutas no tratamento destas alterações posturais características deste transtorno (hiperlordose lombar, anteriorização da cabeça, diminuição da cifose torácica), além de estimular busca de novos meios de intervenções com o enfoque nos diferentes estágios da doença. / Introduction: Schizophrenia is a severe mental disorder that leads to progressive deterioration that more recently has been described as a systemic disease with progressive changes in the brain and body, and partly because of this, associated with lower life expectancy, in average 20 years less than the not affected general population. Although not specific, it has already been determined different inflammatory and oxidative markers involved, however little has been described about systemic markers of schizophrenia. Clinical experience shows that chronic patients are presenting skin, muscles, and posture different from the non patients, but there are no records of studies seeking to study the posture, even though we know that it depends on the interaction of many systems, including the inflammatory state of the subject. Objective: To define patterns of posture within basic parameters (PCL, PRD, PE) in schizophrenic patients and correlate with illness stage and with inflammatory markers of immediate (CRP) and delayed (vWF) response. Method: Cross-sectional study of a convenience sample approved by the ethics committee of the Hospital de Clinicas de Porto Alegre (HCPA). Forty patients with stabilized schizophrenia were recruited in the clinic treatment of HCPA, and analyzed according to 2 subgroups of the stage of the illness (early stage n = 15 less than 10 years from the 1st outbreak, late stage n = 25 10 years or more from the 1st outbreak). The control group (n = 26) were recruited through a social network (Facebook ®). All subjects underwent photogrammetry by SAPO ® method to evaluate posture. The level of significance was set at 5% for all variables and the analyzes were performed using the Statistical Package for the Social Sciences (SPSS, version 18.0). Results: there was no loss, in relation to posture the initial stage group showed 15 angles with significant differences when compared with the reference values, while the late stage group only 7 angles were significant, as compared with the control group only 6angles were significant in a total of 19, the inflammatory markers (CRP and vWF) were not significant in comparison to the initial and late stages of the disease, but the CRP showed correlation with disease severity and the vWF with a postural angle of the protrusion of the head. The pain variable also showed correlation with 5 postural angles, 2 of the column and 3 of the lower limbs, besides the late stage group had a higher prevalence of pain when compared to the early stage. Conclusions: There is a common postural pattern in schizophrenia characterized primarily by protrusion of the head, hyperlordosis and scoliosis, and that early in the illness is more aggravating and in the later stage it stabilizes. And it can be explained by two phenomena: weight gain and muscle weakness. Additionally, we have the influence of pain and inflammatory factors, where the CRP was associated with disease severity but not the posture, but the vWF and the pain were associated with postural angles demonstrating their influences in this disease. This suggests that there is increased pathological activity in the onset of the disease, not only in the brain but also in the body including muscles and adipose tissue. The hyperlordosis appears to be most strongly associated with obesity and adipose tissue, whereas the length of the head and scoliosis more to changes in muscles. All together these findings, if confirmed in larger and more heterogeneous samples will help us in future conduct in treating these postural changes characteristic of this disorder (lumbar hyperlordosis, forward head posture, decreased thoracic kyphosis), besides stimulating the search for new means of interventions with the focus on different stages of the disease.

Introdução: A esquizofrenia é um transtorno mental grave, que com frequência leva a uma deterioração progressiva, que mais recentemente tem sido descrita como uma doença sistêmica, com alterações progressivas cerebrais e corporais, e em parte por causa disto, associada a menor expectativa de vida, em média 20 anos menos que a população geral não afetada. Apesar de não terem especificidade, já foram determinados diferentes marcadores inflamatórios e oxidativos envolvidos; em contrapartida, pouco tem sido descrito sobre marcadores sistêmicos da esquizofrenia. A experiência clínica mostra que os pacientes crônicos vão ficando com pele, musculatura, e postura diferente dos não doentes, porém não há registro de estudo buscando estudar a postura, apesar de sabermos que a mesma depende da interação de inúmeros sistemas, incluindo o estado inflamatório do sujeito. Objetivo: Definir padrões de postura dentro de parâmetros básicos (postura cifolordótica - PCL, postura relaxada ou desleixada - PRD, postura escoliótica - PE) em pacientes esquizofrênicos e correlacionar com fase de doença (inicial-tardia) e com marcadores de resposta inflamatória imediata (proteína C reativa - PCR) e tardia (fator de von Willenbrand - FvW). Método: Estudo transversal, por amostra de conveniência, aprovado pelo comitê de ética do Hospital de Clínicas de Porto Alegre (número 110083 HCPA). Foram recrutados 40 indivíduos com diagnóstico CID-10 e DSMIVTR de esquizofrenia, forma estabilizada, em tratamento ambulatorial no HCPA. Os pacientes foram subdivididos em 2 subgrupos, de acordo com o estágio da doença (estágio inicial n=15 menos de 10 anos do 1° surto, estágio tardio n=25 10 anos ou mais do 1° surto). O grupo controle (n=26) foi recrutado através de uma rede social (Facebook®). Todos os indivíduos foram submetidos à biofotogrametria pelo método SAPO® para avaliar a postura. O nível de significância adotado foi de 5% para todas as variáveis e as análises foram realizadas no programa Statistical Package for the Social Sciences (SPSS, versão 18.0). Resultados: Não houve perdas; em relação à postura o grupo estágio inicial apresentou 15 ângulos com diferenças significativas quando comparado aos valores de referência, enquanto no grupo estágio tardio apenas 7 ângulos foram significativos; já na comparação com o grupo controle, apenas 6 ângulos foram significativos num total de 19. Os marcadores inflamatórios (PCR e FvW) não foram significativos em comparação aos estágios inicial e tardio da doença, porém a PCR apresentou correlação com a gravidade da doença e o FvW com um ângulo postural da protusão da cabeça. A variável dor também apresentou correlação com 5 ângulos posturais, 2 da coluna e 3 dos membros inferiores; além disso o grupo estágio tardio teve maior prevalência de dor quando comparado ao estagio inicial. Conclusões: Existe um padrão postural comum na esquizofrenia caracterizado principalmente por protusão da cabeça, hiperlordose e escoliose, sendo que no início da doença é mais agravante e no estágio mais tardio se estabiliza. Isto pode ser explicado por dois fenômenos: aumento de peso e o enfraquecimento muscular. Adicionalmente, temos a influência da dor e dos fatores inflamatórios, onde a PCR se associou à gravidade da doença, mas não à postura; já o FvW e a dor se associaram aos ângulos posturais, demonstrando suas influências nessa doença. Isto sugere que existe uma maior atividade patológica no início da doença, não só no cérebro, mas também no organismo, incluindo músculos e tecido adiposo. A hiperlordose parece estar mais associada à obesidade e tecido adiposo, enquanto que a extensão da cabeça e escoliose mais a alterações de musculatura. Todos estes achados juntos, se confirmados em amostras maiores e mais heterogêneas, irão nos auxiliar em futuras condutas no tratamento destas alterações posturais características deste transtorno (hiperlordose lombar, anteriorização da cabeça, diminuição da cifose torácica), além de estimular busca de novos meios de intervenções com o enfoque nos diferentes estágios da doença. / Introduction: Schizophrenia is a severe mental disorder that leads to progressive deterioration that more recently has been described as a systemic disease with progressive changes in the brain and body, and partly because of this, associated with lower life expectancy, in average 20 years less than the not affected general population. Although not specific, it has already been determined different inflammatory and oxidative markers involved, however little has been described about systemic markers of schizophrenia. Clinical experience shows that chronic patients are presenting skin, muscles, and posture different from the non patients, but there are no records of studies seeking to study the posture, even though we know that it depends on the interaction of many systems, including the inflammatory state of the subject. Objective: To define patterns of posture within basic parameters (PCL, PRD, PE) in schizophrenic patients and correlate with illness stage and with inflammatory markers of immediate (CRP) and delayed (vWF) response. Method: Cross-sectional study of a convenience sample approved by the ethics committee of the Hospital de Clinicas de Porto Alegre (HCPA). Forty patients with stabilized schizophrenia were recruited in the clinic treatment of HCPA, and analyzed according to 2 subgroups of the stage of the illness (early stage n = 15 less than 10 years from the 1st outbreak, late stage n = 25 10 years or more from the 1st outbreak). The control group (n = 26) were recruited through a social network (Facebook ®). All subjects underwent photogrammetry by SAPO ® method to evaluate posture. The level of significance was set at 5% for all variables and the analyzes were performed using the Statistical Package for the Social Sciences (SPSS, version 18.0). Results: there was no loss, in relation to posture the initial stage group showed 15 angles with significant differences when compared with the reference values, while the late stage group only 7 angles were significant, as compared with the control group only 6angles were significant in a total of 19, the inflammatory markers (CRP and vWF) were not significant in comparison to the initial and late stages of the disease, but the CRP showed correlation with disease severity and the vWF with a postural angle of the protrusion of the head. The pain variable also showed correlation with 5 postural angles, 2 of the column and 3 of the lower limbs, besides the late stage group had a higher prevalence of pain when compared to the early stage. Conclusions: There is a common postural pattern in schizophrenia characterized primarily by protrusion of the head, hyperlordosis and scoliosis, and that early in the illness is more aggravating and in the later stage it stabilizes. And it can be explained by two phenomena: weight gain and muscle weakness. Additionally, we have the influence of pain and inflammatory factors, where the CRP was associated with disease severity but not the posture, but the vWF and the pain were associated with postural angles demonstrating their influences in this disease. This suggests that there is increased pathological activity in the onset of the disease, not only in the brain but also in the body including muscles and adipose tissue. The hyperlordosis appears to be most strongly associated with obesity and adipose tissue, whereas the length of the head and scoliosis more to changes in muscles. All together these findings, if confirmed in larger and more heterogeneous samples will help us in future conduct in treating these postural changes characteristic of this disorder (lumbar hyperlordosis, forward head posture, decreased thoracic kyphosis), besides stimulating the search for new means of interventions with the focus on different stages of the disease.

ADAMTS13 is a metalloprotease that regulates the length, and thus, the platelet-capturing capacity of von Willebrand factor. The regulation of ADAMTS13 activity remains poorly understood. Numerous circulating proteases cleave ADAMTS13 in vitro, impairing its activity, but the physiological significance of this mechanism remains unknown. Two commonly cleaved regions within ADAMTS13 were identified and mutants were developed: two with one of each region mutated (T4L and T8L mutants), one with both regions mutated (T4L/T8L or “double” mutant), and one with an additional elastase site mutated (T4L/T8L + I380G). This work characterizes the mutants’ resistance to proteolysis and compares the activity of the double mutant to wild-type ADAMTS13 (WT). Each mutant and WT was incubated with purified coagulation and neutrophil proteases, activated neutrophils, or added to plasma before initiating coagulation with or without tissue plasminogen activator. Cleavage patterns were visualized with western blot. FRETS-VWF73 and microfluidic flow assays were used to compare WT and mutant activity. Coagulation proteases cleave both predicted sites within WT, and the double mutant exhibits near complete resistance to cleavage over 3 hours. Resistance to degradation by neutrophil proteases is prolonged in the double mutant, but additional cleavage sites are present. Elastase cleavage is prevented in the T4L/T8L + I380G mutant. In plasma, WT is degraded upon initiating coagulation and subsequent fibrinolysis, which is prevented in the double mutant. WT is also degraded in the presence of activated neutrophils, and the double and T4L/T8L + I380G mutants exhibit improved but incomplete resistance. Finally, the mutants exhibit similar activity to WT using FRETS-VWF73 and the microfluidic assay. This work validates the location of two protease-sensitive regions within ADAMTS13 and confirms the resistance of the double mutant to coagulation proteases in vitro. Future work will complete the activity analysis, and compare the mutants’ therapeutic efficacy to WT in vivo. / Thesis / Master of Science (MSc) / Current drugs used to dissolve blood clots can cause major bleeding. Therefore, safer treatments need to be developed. An important step in the clotting pathway is platelet accumulation in the injured vessel. Platelets stick to string-like protein, von Willebrand Factor (VWF), and ADAMTS13 is a protein that regulates this by cutting VWF strings. ADAMTS13 shows promise as a treatment for clots without causing bleeding, but it is unclear how its activity is controlled. ADAMTS13 can be degraded by other proteins, however the importance of this process in the body is unknown. This work characterizes a degradation-resistant ADAMTS13 mutant, which may be used to study whether ADAMTS13 degradation reduces its therapeutic effectiveness. The mutant has normal VWF-cutting activity, is resistant to degradation by clotting proteins, and is partially resistant to proteins released by neutrophils, an important immune cell in clotting. Future studies will investigate its effectiveness at treating clots in animals.

Background: Chronic enteropathies (CE) predispose dogs to thromboembolic disease, but the underlying mechanisms are poorly understood. Humans with CE have decreased activity of a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13), a von Willebrand factor (vWF) cleaving enzyme, and increased circulating vWF. The primary aim of this study is to assess plasma ADAMTS13 activity, vWF antigen (vWF:Ag) concentration, and vWF collagen binding activity (vWF:CBA) in dogs with CE. Hypotheses: Dogs with CE have reduced plasma ADAMTS13 activity, increased vWF:Ag, and increased vWF:CBA compared to healthy control dogs. Animals: Twenty privately-owned dogs with CE and 40 healthy dogs were recruited from a specialty hospital population. Methods: Prospective observational study. Dogs were confirmed to have CE using histopathology. ADAMTS13 activity was measured using a commercially available ELISA kit (DiapharmaⓇ) in 20 dogs with CE and 40 healthy control dogs. Plasma vWF:Ag was assessed in 20 dogs with CE and 20 healthy control dogs. Plasma vWF:CBA was assessed in 19 dogs with CE and 20 healthy control dogs. For statistical analysis, an unpaired t-test was used for normally distributed data and Wilcoxon rank sum was used for non-Gaussian distribution. Significance was set at P < 0.05. Results: Plasma ADAMTS13 activity and vWF:Ag were not significantly different compared to healthy dogs (P = 0.07, P = 0.16, respectively). Plasma vWF:CBA was significantly decreased compared to healthy dogs (P = 0.007). Conclusions and clinical relevance: Plasma ADAMTS13 activity was not significantly different in dogs with CE compared to healthy dogs and is unlikely to be the primary mechanism for hypercoagulability associated with CE. Forty-three percent of CE dogs with hypoalbuminemia had ADAMTS13 activity below reference interval. Further studies are warranted to evaluate ADAMTS13 activity in a subset of dogs with CE, including those with protein losing enteropathy and thromboembolism. / Master of Science / Background: Dogs with chronic gastrointestinal disease, or chronic enteropathies (CE), often have abnormal blood clotting which predisposes to thrombosis. The mechanisms leading to this abnormal blood clotting are poorly understood. Humans with CE also suffer from abnormal blood clotting and thrombosis. Decreased activity of a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13 (ADAMTS13), a von Willebrand factor (vWF) cleaving enzyme, and increased circulating vWF have been reported as contributors to this abnormal blood clotting in people. Von Willebrand factor is a protein that allows platelets to stick together and adhere to damaged tissue to facilitate blood clot formation. The concentration of vWF can be measured using vWF antigen (vWF:Ag) and the activity of vWF can be measured using vWF collagen binding activity (vWF:CBA). Hypothesis: Dogs with CE have reduced plasma ADAMTS13 activity, increased vWF:Ag, and increased vWF:CBA compared to healthy control dogs. Animals: Twenty privately-owned dogs with CE and 40 healthy dogs were recruited from a hospital population. Methods: Prospective observational study. Plasma ADAMTS13 activity, vWF:Ag and vWF:CBA were assessed in dogs with CE and compared to healthy control dogs. Results: There was no difference in plasma ADAMTS13 activity and vWF:Ag between the two groups. The mean vWF:CBA was significantly lower in CE dogs compared to healthy dogs. Conclusions and clinical relevance: Reduced ADAMTS13 activity is unlikely to be the primary mechanism for abnormal blood clotting in dogs with CE.

Studies demonstrated ADAMTS13 possesses unique properties with a mystifying regulatory mechanism. ADAMTS13’s role is in its proteolytic function to its VWF. The disparity in the hemostatic balance between ADAMTS13 activity and the distribution of VWF multimers could result in the bleeding disorder Von Willebrand Disease (VWD) or the thrombotic disorder thrombotic thrombocytopenic purpura (TTP). ADAMTS13 is constitutively secreted as an active protease, yet VWF retains its capacity to recruit platelets. This ability makes ADAMTS13 an enigmatic protease with an unknown regulatory mechanism. Currently, the postulated regulatory mechanism of ADAMTS13 is in its open/closed conformation, yet ADAMTS13 activity is retained in both forms. Literature showed that few proteases are capable of degrading ADAMTS13 in-vitro. We hypothesize that the partial degradation of ADAMTS13 regulates its activity, thereby stabilizing VWF and promoting thrombosis. The goals of this project were to develop and optimize in-vitro plasma BioID to identify novel interactions to ADAMTS13, validate novel interactions, identify proteases capable of degrading ADAMTS13 and their proteolytic sites, and develop protease-resistant ADAMTS13 mutants as novel therapeutics to thrombotic disorders. We optimized the BioID technique to be used in-vitro in plasma, to study novel interactions with ADAMTS13. Our results identified novel potential interactions with vitronectin or plasminogen. Validation studies disregarded vitronectin’s interaction and confirmed plasminogen’s interaction through the CUB and Kringle domains in a lysine-dependent manner. Further, the list of proteases capable of degrading ADAMTS13 was expanded to include FXIa and neutrophil-derived proteases including Cathepsin G, elastase, and hPR3. Activated neutrophils played a stronger role than coagulation proteases in degrading ADAMTS13 in vivo, while also demonstrating that elastase is a more potent regulator. Proteolytically degraded sites on ADAMTS13 were identified and proteolytic-resistant ADAMTS13 mutants were produced accordingly, which we aim to be utilized as a novel therapeutic to thrombotic disorders. / Thesis / Doctor of Philosophy (Medical Science)