Dissertation (Ph.D.)--University of South Florida, 2007. / Includes vita. Includes bibliographical references (leaves 109-141). Also available online.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Some forms of neuropathic pain are maintained by sympathetic fibers, but the underlying mechanisms are poorly understood. Thus, the aim of this study was to investigate the possible involvement of the TRPA1 receptor as well as the role of the sympathetic nervous system (involved in sympathetically maintained neuropathic pain) in a model of neuropathic pain induced by chronic sciatic nerve constriction injury (CCI) in mice. The systemic injection of the selective TRPA1 antagonist HC-030031 reversed both mechanical and cold allodynia induced by chronic sciatic nerve constriction injury. Nerve injury also sensitizes mice to the nociception induced by the intraplantar injection of a low dose of the TRPA1 agonist allyl isothiocyanate without changing TRPA1 immunoreactivity in the injected paw. Furthermore, the chemical sympathectomy produced by guanethidine largely prevented CCI-induced mechanical and cold allodynia. CCI also induced a norepinephrine-trigged nociception that was inhibited by α-adrenoceptor antagonism and norepinephrine transporter and monoamine oxidase inhibition. Finally, the peripheral injection of HC-030031 also largely reduced CCI-induced nociception by norepinephrine and mechanical or cold allodynia. Taken together, the present findings reveal the critical role of TRPA1 in mechanical and cold hypersensitivity as well as in hypersensitivity to norepinephrine following nerve injury. This article presents the role of TRPA1 receptor on the sympathetically-maintained nociception induced by nerve injury in mice. Our results suggest that TRPA1 antagonists may be useful to treat neuropathic patients that present sympathetically maintained pain. / Algumas formas de dor neuropática são mantidos por fibras simpáticas, contudo os mecanismos subjacentes são pouco compreendidos. Assim, o objetivo deste estudo foi investigar o possível envolvimento do receptor TRPA1, bem como o papel do sistema nervoso simpático num modelo de dor neuropática induzido pela constrição crônica do nervo ciático (CCI) em camundongos. A administração sistêmica do antagonista seletivo do receptor TRPA1, HC-030031, reverteu a alodinia mecânica e ao frio induzida pela CCI. A injeção de uma baixa dose do agonista do receptor TRPA1, isotiocianato de alila, induziu uma resposta nociceptiva nos animais que sofreram a lesão no nervo, sem alterar a imunorreatividade do TRPA1 na pata injetada. Além disso, a simpatectomia química produzida pela guanetidina amplamente previniu a alodínia mecânica e ao frio induzida pela CCI. Ainda, a injeção intraplantar de norepinefrina induziu nocicepção espontânea, a qual foi reduzida pelo antagonista α-adrenérgico, inibidor do transportador de norepinefrina e inibidor da enzima monoamina oxidase. Finalmente, a administração periférica do HC-030031 reduziu a nocicepção espontânea induzida pela norepinefrina e também a alodínia mecânica e térmica de animais neuropáticos. Assim, nossos resultados revelam o papel crítico do receptor TRPA1 na alodínia mecânica e ao frio, bem como na hipersensibilidade à norepinefrina após lesão do nervo em um modelo de dor neuropática mantida pelo simpático. Dessa forma, o receptor TRPA1 poderá ser um alvo para o desenvolvimento de novos tratamentos para esse tipo de dor.

Made available in DSpace on 2015-07-22T20:49:28Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-06-29 / Objetivo: Avaliar a atencao visual de criancas com migranea e comparar aos controles. Metodos: Foram avaliadas 30 criancas com migranea com (n=5) e sem aura (n=25), pelos criterios da gInternational Headache Society (ICHD-II, 2004) h, 15 delas do sexo feminino, faixa etaria entre 8 e 12 anos e 11 meses (media=10,8 anos, SD=1,5), media de 5,8 dias de cefaleia/mes (SD=3,2), confirmadas em diario de cefaleia, e 30 controles sem cefaleia, 14 do sexo feminino, idade media de 9,9 anos (SD=1,3). Testes de atencao visual utilizados: gTrail Making Test h A/B (avaliado tempo de execucao em segundos); Teste de Cancelamento de Letras (avaliados tempo de execucao em segundos e o numero de erros) e o teste computadorizado TAVIS-3 (Teste de Atencao Visual . 3a edicao), avaliando tempo de reacao (TR), erros por omissao (EO) e por acao (EA) atraves de subtestes de atencao seletiva (tarefa 1), alternada (tarefa 2) e sustentada (tarefa 3). No momento da avaliacao, as criancas com migranea estavam sem cefaleia ha pelo menos dois dias. Criterios de exclusao: alteracoes do exame neurologico, doencas sistemicas, epilepsia, trauma craniano, disturbios psiquiatricos, disturbios de sono, disturbios de aprendizado, uso de medicacoes, incluindo profilaticas para migranea, drogas ou alcool, e Quociente de Inteligencia (QI) total < 80, avaliado pela Escala Wechsler de Inteligencia 3a edicao (WISC-III). Analise estatistica: testes t de Student e Mann-Whitney e Nivel de Confianca de 95% (p.0,05). Resultados: Criancas com migranea apresentaram desempenho inferior aos controles nos testes: gTrail Making Test h A (p=0,03) e B (p=0,001) e mais erros por acao (EA) nas tarefas 1 (p=0,032), 2 (p=0,015) e na soma de erros por acao das tres tarefas (EA TOTAL) do TAVIS-3, apresentando dificuldades nas tres modalidades atencionais avaliadas, mas, principalmente, em atencao seletiva e alternada. Discussao: A atencao e uma funcao neurologica complexa envolvendo estruturas anatomicas como tronco cerebral, cortex, sistema limbico e neurotransmissores, como a dopamina e noradrenalina. Os resultados desse estudo permitem supor que aspectos neuroquimicos envolvidos tanto na fisiopatologia da migranea, como nos mecanismos atencionais, predisponham essas criancas a deficits de atencao visual. Conclusao: criancas com migranea apresentaram pior desempenho nos testes de atencao visual quando comparadas com criancas sem cefaleia. / Objective: To evaluate the visual attention of children with migraine and compare to the headache free control group. Method: 30 children with migraine, with (n=5) and without aura (n=25), which fulfill the gInternational Headache Society (ICHD-II, 2004) h criteria, 15 female, ages 8 to 12 years and 11 months (average=10.8 }1.5 years), with an average of 5.8 } 3.2 attacks/month, lasting up to 24 hours, and 30 control subjects without headache, 14 female, average age 9,9 }1.3 years. Method of assessment of visual attention: Trail Making Test (TMT) A/B, Letter Cancellation Test, and the computerized test Visual Attention Test . 3rd edition (TAVIS-3) which evaluates reaction time, omission and action errors through subtests of selective (task1), alternate (task2), and sustained attention (task3). Migraine group were evaluated after two days without headache. Exclusion criteria: other systemic diseases, altered neurological examination, psychiatric disorders, learning disorders, epilepsy, head trauma, use of medication (including migraine prophylaxis), and an Intelligence Quotient (IQ) < 80, measured by Wechsler Intelligence Scale 3rd edition. Statistical analysis: Student fs t test and Mann-Whitney fs test, Confidence Interval of 95% (p.0.05). Results: Children with migraine had inferior performance compared to the control group on TMT A (p=0.03) and B (p=0.001), and more action errors on tasks 1 (p=0.032) and 2 (p=0.015) of TAVIS-3, presenting difficulty with selective and alternate attention. Discussion: Attention is a complex neurologic function which depends on structures such as the brain stem, cerebral cortex, and the limbic system and on neurotransmitters such as dopamine and noradrenaline. Conclusion: The neurochemical aspects involved in the physiopathology of migraine and attention mechanisms probably predispose these children to visual attention deficits. / TEDE / BV UNIFESP: Teses e dissertações

Made available in DSpace on 2014-06-11T19:32:06Z (GMT). No. of bitstreams: 0 Previous issue date: 2009-08-21Bitstream added on 2014-06-13T20:03:01Z : No. of bitstreams: 1 bernabe_dg_dr_araca.pdf: 658277 bytes, checksum: bf79196ee6bff2396cbef9ab03e7fc4b (MD5) / Introdução: Estudos têm mostrado que o estresse pode influenciar o desenvolvimento e a progressão do câncer. Entretanto, pouco se sabe sobre a interferência de fatores neurohormonais relacionados ao estresse, como o cortisol e a catecolamina norepinefrina (NE), sobre o carcinoma espinocelular (CEC) de boca. Proposição: Neste estudo, nós investigamos se pacientes portadores de CEC de boca apresentam alterações nos níveis matinais de cortisol plasmático e salivar, da quimiocina CXCL12 plasmática e de óxido nítrico (NO) salivar. Além disso, investigamos os efeitos da NE, isoproterenol (um agonista ß-adrenérgico) e cortisol sobre linhagens celulares derivadas de CEC de boca. Pacientes e métodos: Foram avaliados 34 pacientes com CEC de boca, 17 pacientes com CEC de orofaringe, 17 portadores de leucoplasia bucal, 27 indivíduos expostos ao tabagismo e/ou alcoolismo e 25 voluntários saudáveis. Os níveis salivares e plasmáticos de cortisol e os níveis plasmáticos de CXCL12 foram analisados por ELISA. A dosagem de NO salivar foi feita pelo método de Griess. A associação com variáveis clínicas como idade, sexo, consumo de cigarro e álcool, estádio clínico e ciência do diagnóstico foi investigada. Os efeitos da NE, do isoproterenol e do cortisol sobre a expressão de Interleucina-6 (IL-6) e NO foram investigados nas células derivadas de CEC de boca SCC9 e SCC25. A expressão de IL-6 pelas linhagens celulares foi analisada por PCR em tempo real e a produção da proteína no sobrenadante celular por ELISA. A dosagem de NO no sobrenadante foi feita pelo método de Griess. Os efeitos dos hormônios do estresse sobre a proliferação celular das células SCC9 foram avaliados por MTT. A expressão de receptores adrenérgicos ß1 e ß2 foi analisada nas linhagens SCC9 e SCC25 e em amostras humanas de CEC de boca, leucoplasia e mucosa bucal normal por PCR em tempo... / Background: Investigations had showed that stress can influence the development and progression of cancer. However little is known about the interference of stressrelated neurohormones factors on the oral squamous cell carcinoma (OSCC). Purpose: In this study we investigated whether patients with OSCC show changes in levels of morning salivary and serum cortisol, serum CXCL12 and salivary nitric oxide (NO). Moreover we evaluated the effects of stress hormones on cell lines derived from OSCC. Patients and methods: The serum levels of cortisol and CXCL12 and the levels of salivary cortisol and NO (form of NO2) were studied in thirty-four patients with OSCC, and compared with seventeen patients with oropharynx SCC, seventeen patients with oral leukoplakia, twenty-seven individuals exposed to cigarette smoke and / or alcohol and twenty-five healthy volunteers. The association with clinical data such as age, sex, consumption of cigarettes and alcohol, clinical stage and the consciousness of diagnosis was investigated. The effects of stress-related mediators including norepinefrine (NE), isoproterenol and cortisol on the production of Interleukine-6 (IL-6) and nitric oxide were studied in the cells derived from OSCC SCC9 and SCC25. The IL-6 expression was evaluated by real time PCR and the protein concentration in supernatants using ELISA. The concentration of NO in culture supernatants was measured with Griess reagent. The effects of hormones on cell proliferation of SCC9 cells was evaluated by MTT. The expression of ßadrenergic receptors was evaluated by real time PCR in SCC9 and SCC25 cells and in 20 samples of OSCC, 17 of leukoplakia and 15 of normal oral mucosa samples. Results: The serum and salivary levels of cortisol were significantly higher in patients with OSCC compared with patients with leukoplakia, smokers and / or drinkers and... (Complete abstract click electronic access below)

Les effets des catécholamines en réanimation sont peu prédictibles. La variabilité interindividuelle des observations est d’origine multifactorielle dont des facteurs pharmacocinétiques et pharmacodynamiques, dépendant de caractéristiques constitutionnelles et acquises de chaque individu. Les posologies de l’adrénaline et de la noradrénaline chez l’enfant sont extrapolées des données adultes. Pourtant l’âge est une source de grande variabilité liée au développement. Un modèle pharmaco- statistique de l’adrénaline et de la noradrénaline a été établi chez l’enfant en insuffisance circulatoire aigüe. Il a permis d’identifier des facteurs de variabilité entre les individus ainsi que de proposer des schémas de prescription des deux molécules en fonction de l’effet souhaité et des caractéristiques de l’enfant. La pharmacocinétique de l’adrénaline chez 39 enfants en prévention du syndrome de bas débit cardiaque postopératoire suivait un modèle monocompartimental. La clairance augmentait avec le poids selon le principe de l’allométrie. Les augmentations résultantes de la fréquence cardiaque et de la pression artérielle moyenne suivaient un modèle d’effet direct Emax. Elles étaient influencées par l’âge et la gravité des patients. Les augmentations de glycémie et lactatémie suivaient un modèle d’effet indirect. Pour la noradrénaline, chez 38 enfants atteints d’hypotension artérielle systémique, la pharmacocinétique était mono-compartimentale. La clairance était influencée par le poids (allométrie). L’augmentation induite de la pression artérielle moyenne suivait un modèle direct Emax. Elle était fonction de l’âge et de la gravité des patients. Les posologies de l’adrénaline et de la noradrénaline chez l’enfant devraient tenir compte du poids, de l’âge et de la gravité du patient : plus jeune est l’enfant et plus grave est son état, plus la posologie doit être élevée pour satisfaire les objectifs hémodynamiques. / The effects of catecholamines are difficult to predict. The between-subject variability observed in clinical setting is multifactorial including constitutional and acquired characteristics of each individual. Epinephrine and norepinephrine dosages are usually extrapolated from adult data. Yet, age is a source of high variability due to development- related phenomena. A population model of epinephrine or norepinephrine was developed in haemodynamically critically ill children. This allowed to identify between-subject variability factors as well as to propose individualized dosage regimens of these two catecholamines according to the desired effect and child characteristics. Epinephrine pharmacokinetics in 39 children at high risk of postoperative low cardiac output syndrome followed a one-compartment model. Clearance increased with bodyweight according to the allometric rule. The resulting increases in heart rate and mean arterial pressure followed a direct Emax model. These were related to age and illness severity. A turn-over model described the increases in blood glucose and lactate. Norepinephrine pharmacokinetics in 38 hypotensive critically ill children followed a one compartment model. Clearance increased with bodyweight (allometry). The resulting increase in mean arterial pressure followed a direct Emax model. This was a function of age and illness severity. The dosage of epinephrine and norepinephrine in children should take into account the bodyweight, age and illness severity of the patient: the younger the child and the more serious the condition, the higher the dosage in order to meet the haemodynamic goals.

Práca sa zaoberá in silico dizajnom a validáciou peptidov založených na konotoxíne - MrIA, izolovaného z morských slimákov druhu Conus marmoreus a možnosti využitia týchto peptidov v liečbe neuroblastómu pomocou cielenia norepinefrínového transportéru. Päť peptidov založených na tomto konotoxíne bolo simulovaných pomocou simulácii molekulárnej dynamiky, ich trajektórie boli analyzované pre zistenie vlastností týchto peptidov. Dva homologické modely ľudského norepinefrínového transportéru boli vytvorené pre analýzu väzobných vlastností peptidov založených na konotoxíne ku norepinefrínovému transportéru. Peptidy boli následne syntetizované a použité na pokrytie apoferitínových nanočastíc s elipticínom uväzneným vnútri apoferitínu. Vytvorené peptidy a nanočastice boli ďalej skúmané pre objasnenie ich fyzikálo-chemických vlastností. Interakcie a cytotoxicita boli skúmané aplokáciou nanočastíc na bunky neuroblastómu a epitelu. Z in silico a in vitro analýz vyšiel YKL-6 peptid ako najlepší kandidát na ďalší výskum.

L'attention est une fonction au cœur de la cognition qui, à tout moment, nous permet de sélectionner les informations pertinentes à traiter, tout en ignorant les autres. Cette sélection de l’information qui s’opère à la fois dans l'espace et dans le temps résulte de l’intégration des informations sensorielles et d’un contrôle de "haut niveau" en fonction de nos buts. Cette fonction dépend d’un réseau cérébral incluant le système fronto-pariétal et est sous l’influence de différents neuromodulateurs, en particulier la noradrénaline, dont l’action reste encore mal connue. Mon travail de thèse consistait à comprendre le rôle de la noradrénaline sur les processus attentionnels. Mes objectifs étaient d’une part de vérifier notre hypothèse selon laquelle la noradrénaline modulerait les différentes facettes de l’attention (attention spatiale et attention soutenue) et d’autre part d’élucider les mécanismes d’action par lesquelles la noradrénaline exercerait ces effets. Pour répondre à ces questions, nous avons testé l’impact d’une augmentation de la transmission noradrénergique (administration intramusculaire d'atomoxétine) chez le singe, dans des tâches comportementales nécessitant une sélection de l’information visuelle soit dans l’espace (tâche d'attention avec indice et exploration spontanée d'images) soit au cours du temps (tâche de discrimination go/nogo). Nos résultats démontrent que l’atomoxétine facilite les processus attentionnels à la fois dans l’espace et au cours du temps. Dans l’espace, l’atomoxétine module l’orientation de l’attention visuo-spatiale en fonction du contexte, en ajustant le taux d’accumulation sensorielle ou l’impact de la saillance des images sur l’orientation de l’attention. Au cours du temps, l’atomoxétine ajuste la relation entre la sensibilité à discriminer la cible parmi des distracteurs et le biais de réponse des animaux. En résumé, mes résultats démontrent que la noradrénaline influence les deux facettes, spatiale et temporelle de l’attention et suggèrent une action via un ajustement des processus de traitement de l’information sensorielle et un ajustement du contrôle de l’attention au contexte / Attention is a function at the heart of cognition that, at any given moment, enables us to select some information for further processing, while setting aside others. This selection of information that operates both in space and time, results from the integration of sensory information and higher-level control according to our goals. This function depends on a cerebral network including the fronto-parietal system. It is also under the influence of different neuromodulators, in particular norepinephrine, the action of which is still poorly understood.The aim of my PhD work was to understand the role of norepinephrine on attentional processes. My objectives were, on the one hand, to test our hypothesis that norepinephrine is capable of acting on the different facets of attention (spatial attention and sustained attention) and, on the other hand, to elucidate the mechanisms of action by which noradrenaline exerts its action. To answer these questions, we tested the impact of an increase in noradrenergic transmission (intramuscular administration of atomoxetine) in monkeys, using behavioral tasks requiring a selection of visual information in space (cued attentional task and spontaneous image exploration) or over time (go/nogo discrimination task). Our results demonstrate that atomoxetine facilitates attentional processes both in space and over time. In space, atomoxetine modulates the orientation of visuospatial attention according to the context, adjusting the rate of sensory accumulation or the impact of image saliency on attention orientation. Over time, atomoxetine adjusts the relationship between the sensitivity to discriminate a target among distractors and the animal’s response bias.In summary, my results demonstrate that norepinephrine influences both the spatial and temporal facets of attention and suggests an action through an adjustment of sensory information processing and an adjustment of attention control to the context

Generalized anxiety disorder (GAD) is associated with painful physical symptoms (PPS). These post hoc analyses of previous trial data assessed PPS and their response to duloxetine treatment in GAD patients. Studies 1 and 2 (n = 840) were 9- to 10-week efficacy trials; study 3 (n = 887) was a relapse prevention trial comprising a 26-week open-label treatment phase and a 26-week double-blind, placebo-controlled treatment continuation phase. Mean baseline visual analog scale scores (VAS, 0–100; n = 1727) ranged from 26 to 37 for overall pain, headache, back pain, shoulder pain, interference with daily activities, and time in pain while awake. In studies 1 and 2, improvement on all VAS scores was greater in duloxetine-treated than in placebo-treated patients (p ≤ 0.01). In study 3, pain symptoms worsened in responders switched to placebo compared with those maintained on duloxetine (p ≤ 0.02). In conclusion, duloxetine was efficacious in the short- and long-term treatment of PPS, which are common in GAD patients.

Heart failure is the main cause of death for both men and women in the United States. The only proven treatment for patients with heart failure is heart transplantation. The goal of this research is to create patches of tissue that could mimic the function of the native heart to repair the damaged portions of the heart. In this study, whole porcine hearts were decellularized to create a 3D construct that was recellularized with cardiomyocytes (CM) differentiated from human induced pluripotent stem (IPS) cells. At day 4 of differentiation, IPS-derived CMs were implanted onto cardiac extracellular matrix (cECM) and ten days after recellularization, the cells started to beat spontaneously. After implantation, the progenitor CMs continued to proliferate and populate the cECM. A live/dead assay showed the potential of the cECM as a scaffold suitable for recellularization. Confocal microscopy images were taken to evaluate the organization of the cells within the matrix and the impact of the cECM on the growth and maturation of the CMs. Representative cardiac Troponin T (cTNT) and vimentin immunostaining images of CMs derived from iPSCs, on cECM and on standard cell culture plates showed that the cECM allowed the cells to organize and form fibrils with the fibroblasts, compared with CMs cultured in regular culture plates. The timeline of implantation of the cells was a key factor for the development of the heart tissue constructs. Progenitor CMs seeded onto cECM showed better organization and the ability to penetrate 96 µm deep within the collagen fibers and align to them. However, mature CMs seeded onto the matrix showed a disorganized network with very reduced interaction of CMs with fibroblasts, forming two different layers of cells; CMs on top of fibroblasts. In addition, the depth of penetration of the mature CMs within the matrix was only 20 µm. To evaluate the impact of the addition of support cells to the CM monolayer cultures, CMs were co-cultured with human umbilical vein endothelial cells (HUVEC) and it was demonstrated that at ratios of 2:1 HUVEC:CM the beating rate of the CMs was improved from 20 to 112 bpm, additionally, the CM monolayer cultures showed a more synchronized beating pace after the addition of HUVECs. Pharmacological stimulation was performed on CM monolayer cultures using norepinephrine as a stimulator and the results showed that the beating pace of the CMs was improved to 116 bpm after 5 minutes of drug exposure. For future studies, inosculation of the tissue constructs could be performed with the incorporation of membrane proteins to understand the mechanotransduction of the cells. As a preliminary study, the action of dual claudins was evaluated with HUVEC cultures and the results showed the potential of these membrane proteins in the healing of the damaged cell membrane.

No description available.