HOXA9 as a key regulator in glioma initiation, aggressiveness and response to therapy

Tese de Doutoramento em Ciências da Saúde

Satisfação profissional dos Técnicos de Diagnóstico e Terapêutica

Dissertação de mestrado em Gestão de Unidades de Saúde

Ventricular evoked response in patients with hypertrophic obstructive cardiomyopathy treated with DDD pacing

OBJECTIVE: To assess the changes in ventricular evoked responses (VER) produced by the decrease in left ventricular outflow tract gradient (LVOTG) in patients with hypertrophic obstructive cardiomyopathy (HOCM) treated with dual-chamber (DDD) pacing. METHODS: A pulse generator Physios CTM (Biotronik, Germany) was implanted in 9 patients with severe drug-refractory HOCM. After implantation, the following conditions were assessed: 1) Baseline evaluation: different AV delay (ranging from 150ms to 50 ms) were sequentially programmed during 5 to 10 minutes, and the LVOTG (as determined by Doppler echocardiography) and VER recorded; 2) standard evaluation, when the best AV delay (resulting in the lowest LVOTG) programmed at the initial evaluation was maintained so that its effect on VER and LVOTG could be assessed during each chronic pacing evaluation. RESULTS: LVOTG decreased after DDD pacing, with a mean value of 59 ± 24 mmHg after dual chamber pacemaker, which was significantly less than the gradient before pacing (98 + 22mmHg). An AV delay >100ms produced a significantly lower decrease in VER depolarization duration (VER DD) when compared to an AV delay <=100ms. Linear regression analyses showed a significant correlation between the LVOTG values and the magnitude of VER (r=0.69; p<0.05) in the 9 studied patients. CONCLUSION: The telemetry obtained intramyocardial electrogram is a sensitive means to assess left ventricular dynamics in patients with HOCM treated with DDD pacing.

Response of blood pressure to maximum exercise in hypertensive patients under different therapeutic programs

OBJECTIVE: To evaluate the behavior of blood pressure during exercise in patients with hypertension controlled by frontline antihypertension drugs. METHODS: From 979ergometric tests we retrospectively selected 49 hipertensive patients (19 males). The age was 53±12 years old and normal range rest arterial pressure (<=140/90 mmHg) all on pharmacological monotherapy. There were 12 on beta blockers; 14 on calcium antagonists, 13 on diuretics and 10 on angiotensin converting enzyme inhibitor. Abnormal exercise behhavior of blood pressure was diagnosed if anyone of the following criteria was detected: peak systolic pressure above 220 mmHg, raising of systolic pressure > or = 10 mmHg/MET; or increase of diastolic pressure greater than 15 mmHg. RESULTS: Physiologic response of arterial blood pressure occurred in 50% of patients on beta blockers, the best one (p<0.05), in 36% and 31% on calcium antagonists and on diuretics, respectively, and in 20% on angiotensin converting enzyme inhibitor, the later the leastr one (p<0.05). CONCLUSION: Beta-blockers were more effective than calcium antagonists, diuretics and angiotensin-converting enzyme inhibitors in controlling blood pressure during exercise, and angiotensin converting enzyme inhibitors the least effective drugs.

Evaluation of inflammatory response induced by different types of daily contact lenses

Dissertação de mestrado em Genética Molecular

Endothelium-dependent relaxation in response to poly-L-arginine in canine coronary arteries: implications about hyperpolarization as a mechanism of vasodilatation

OBJECTIVE: To study the mechanism by which poly-L-arginine mediates endothelium-dependent relaxation. METHODS: Vascular segments with and without endothelium were suspended in organ chambers filled with control solution maintained at 37ºC and bubbled with 95% O2 / 5% CO2. Used drugs: indomethacin, acetycholine, EGTA, glybenclamide, ouabain, poly-L-arginine, methylene blue, N G-nitro-L-arginine, and verapamil and N G-monomethyl-L-arginine. Prostaglandin F2á and potassium chloride were used to contract the vascular rings. RESULTS: Poly-L-arginine (10-11 to 10-7 M) induced concentration-dependent relaxation in coronary artery segments with endothelium. The relaxation to poly-L-arginine was attenuated by ouabain, but was unaffected by glybenclamide. L-NOARG and oxyhemoglobin caused attenuation, but did not abolish this relaxation. Also, the relaxations was unaffected by methylene blue, verapamil, or the presence of a calcium-free bathing medium. The endothelium-dependent to poly-L-arginine relaxation was abolished only in vessels contracted with potassium chloride (40 mM) in the presence of L-NOARG and indomethacin. CONCLUSION: These experiments indicate that poly-L-arginine induces relaxation independent of nitric oxide.

Immune cell changes in swimmers : response to acute exercise and training

O treino competitivo envolve exercício intenso e prolongado, capaz de modular o número e actividade das células imunitárias. Quando demasiado exigente poderá induzir fadiga e aumentar a susceptibilidade a doenças. Esta dissertação apresenta três estudos desenvolvidos no âmbito da Imunologia do Exercício, considerando a análise da resposta celular imunitária sistémica aguda e crónica ao exercício aplicada em situações reais do treino competitivo de natação, controlando factores passíveis de influenciar esta resposta. Pretendeu-se avaliar a resposta imunitária a uma sessão de treino prolongada e intensa, durante as 24h de recuperação (Estudo 1) e a uma época de treino com sete meses (Estudo 2), e estudar a influência de um macrociclo de treino de quatro meses sobre a resposta imunitária à mesma sessão de treino e período de recuperação (Estudo 3), controlando sexo, fases do ciclo menstrual, maturidade, escalão, especialidade, performance, cargas de treino e sintomas respiratórios superiores (URS). A sessão de treino induziu a diminuição da vigilância imunitária adquirida imediatamente e, pelo menos nas 2h seguintes. Juvenis e seniores recuperaram totalmente 24h depois, mas não os juniores, reforçando a ideia da existência de uma janela aberta para a infecção após exercícios prolongados e intensos e sugerindo uma recuperação menos eficiente para os juniores. No período de treino mais intenso da época observou-se uma imunodepressão e maior prevalência de URS. No final da época, a imunidade inata diminuiu aparentando maior sensibilidade aos efeitos cumulativos da carga de treino, enquanto a imunidade adquirida parece ter recuperado após o taper. O macrociclo de treino atenuou a resposta imunitária à sessão de treino e aumentou o período de janela aberta às infecções (efeitos mais acentuados nos adolescentes). Os resultados evidenciam a importância de controlar alterações imunitárias durante a época competitiva, especialmente em períodos de treino intenso e quando se realizam sessões de treino intensas consecutivas com recuperações inferiores a 24h.

Senescencia de celulas T humanas inducida por tumores: un nuevo mecanismo de evasión de la respuesta inmune. Senescence of human T cells iduced by tumors. A new mechanism of evasion of immune response.

La respuesta antitumoral en individuos con cáncer depende, en gran medida, de células del sistema inmune capaces de reconocer y eliminar las células tumorales. Sin embargo, los tumores tienen la capacidad de evadir la respuesta inmune a través de diversos mecanismos como por ejemplo inducir la muerte de células claves del sistema inmune [1-3]. Previamente nosotros demostramos mediante un modelo in vitro que dependiendo de las condiciones de interacción entre tumor y linfocitos (tiempo y relación numérica), los tumores pueden inducir apoptosis o senescencia de linfocitos T provenientes de donantes sanos. Nuestros resultados son los primeros en demostrar que células tumorales de diversos orígenes pueden inducir senescencia de células T a través de factor/s solubles. También demostramos que a diferencia con los que ocurre in vivo, tanto las células CD4 como las CD8 son susceptibles a adquirir fenotipo de senescencia. Estudiando las implicancias que puede tener la senescencia sobre la funcionalidad de la célula T, observamos que las células T CD4+ y CD8+ senescentes pueden suprimir una respuesta linfoproliferativa. Si bien las células CD8+CD28- han sido identificadas in vivo, nosotros demostramos que células CD4+ CD28- tiene capacidad supresora [4]. En base a estos resultados, nuestra hipótesis es que las células T senescentes inducida por tumores pueden regular nuestro sistema de defensa actuando sobre la respuesta inmune adaptativa y posiblemente sobre la innata y, por consiguiente, postulamos que la senescencia de células T puede ser considerada como otro de los mecanismos de evasión de la respuesta inmunePlanteamos así los siguientes objetivos específicos: -Evaluar como las células T senescentes inducidas por tumores pueden regular la respuesta inmune.-Evaluar la participación de mediadores solubles capaces de regular la senescencia de células T inducida por tumores. En la actualidad existen estrategias inmuno-terapéuticas que avizoran resultados promisorios. Sin embargo, el control de células T inmunosupresoras permanece como unos de los grandes desafíos. Nuestro proyecto proveerá conocimientos sobre un fenómeno muy poco estudiado y por consiguiente muy poco valorado a la hora de diseñar estrategias terapéuticas para la cura del cáncer.

Regulación de la actividad funcional tiroidea e interacción con mediadores de la respuesta inmune. Regulation of thyroid functional activity and interaction with mediators of immune response.

TEMA I. MECANISMOS NO CLÁSICOS EN EL CONTROL DE LA BIOSÍNTESIS DE HORMONAS TIROIDEAS. El objetivo general de este aspecto del proyecto es el estudio de los mecanismos bioquímicos y moleculares que regulan la hormonogénesis tiroidea en diferentes condiciones funcionales. En proyectos anteriores hemos demostrado que la endotoxina bacteriana lipopolisacárido (LPS) y el oxido nítrico (NO) inducen modificaciones en la biosíntesis de hormonas tiroideas. En base a estos resultados se propone investigar el mecanismo responsable de la estimulación de la captación de ioduro y la expresión de NIS ejercida por LPS y los factores que regulan la producción de NO en la célula tiroidea. Por otra parte se investigarán posibles factores hormonales reguladores de la absorción de ioduro a nivel intestinal y su relación con el eje hipófiso-tiroideo, así como los mecanismos involucrados en la expresión de NIS en enterocitos. Como extensión con aplicación clínica y en base a la experiencia del grupo en el estudio de proteínas que participan en la biosíntesis hormonal tiroidea, se realizará una pesquisa de posibles mutaciones en el transportador de ioduro en pacientes con Hipotiroidismo congénito.TEMA II. ESTUDIO DEL EFECTO DE LAS HORMONAS TIROIDEAS EN LA INICIACIÓN DE LA RESPUESTA INMUNE EN RATÓN. INTERACCIÓN CON GLUCOCORTICOIDES. En nuestro grupo demostramos recientemente un efecto novel de las hormonas tiroideas sobre la maduración/función de células presentadoras de antígenos especializadas: células dendríticas (DC), derivadas de médula ósea de ratón. En este proyecto se propone: 1) profundizar el mecanismo molecular involucrado en el efecto de triiodotironina (T3) sobre DC: rol del receptor de T3-señalamiento intracelular; capacidad de DC tratadas con T3 de estimular la citotoxicidad antígeno-específica; estrategia de vacunación en el tratamiento antitumoral. Por su parte, también previamente demostramos la característica de los glucocorticoides (GC) de interaccionar con el mecanismo de acción de las hormonas tiroideas en la expresión final de los efectos T3-específicos. Considerando el uso terapéutico de los GC en diversos estados clínico-patológicos inmunes, se propone: 2) Estudio del efecto de dexametasona (GC de síntesis) sobre el mecanismo de acción de T3 a nivel de DC.

Participación de los receptores de Angiotensina AT1 de la Amigdala en la respuesta al miedo potenciado. Role of Amygdalar Angiotensin AT1 receptors in response to fear potentiated.

La amígdala se encuentra estrechamente ligada a la generación y modulación de los procesos emocionales. Aunque el complejo de la amígdala generalmente se define por varios grupos distintos de células, los núcleos basolaterales que se conectan con el núcleo central y el núcleo de la estría terminal son los que proyectan a las áreas del sistema nervioso central involucradas en el control de las respuestas autónomas, los procesos cognitivos y la respuesta emocional. Además de los sistemas glutamatérgico, gabaérgico, CRH, Opiodes, CCK entre otros, en estas áreas de la amígdala se encuentran receptores AT1 del Sistema Renina-Angiotensina (SRA) cerebral. Entre las áreas a las que se proyectan estos núcleos se destaca la inervación de núcleos dopaminérgicos a través del área tegmental ventral y su influencia sobre la función del eje hipotálamo hipófiso adrenal por la modulación de la descarga de ACTH a través de la inervación del núcleo hipotalámico paraventricular. También se ha comprobado la colocalización del SRA y receptores AT1 de Angiotensina II en sustancia nigra y cuerpo estriado, sugiriendo un rol clave de SRA en la modulación de la liberación de dopamina central. Existen evidencias, neuroanatómicas, fisiológicas y farmacológicas que indican que la Angiotensina II cerebral es mediadora de las respuestas inducidas por estrés incluyendo la regulación de los sistemas simpático y neuroendócrino. En este proyecto se propone evaluar en ratas wistar la participación de los receptores AT1 de Angiotensina II en la respuesta al miedo potenciado. En este modelo hay una mayor activación de la amígdala y el establecimiento de un estado de ansiedad por la exposición previa a una situación de estrés que desencadena respuestas similares a las encontradas en pacientes con desordenes de ansiedad. Además evaluaremos su posible participación en las conducta de exploración, expresión de la memoria de trabajo y recambio de dopamina en los núcleos del estriado, accumbens y corteza prefrontal.

Synaptic plasticity in mature cultured hippocampal-entorhinal cortex slices : activity-dependent regulation of cAMP response-element binding protein

LTP, synaptic plasticity, hippocampus, organotypic cultures, CREB

Neural dynamics and their relationship to learning behavior in response tot stimulation with a cortical neuroprosthesis

Sensory cortex, neuroprosthetics, brain-machine-interfaces, neurodynamics, learning, perception, embodied cognition

Vascular Response of Ruthenium Tetraamines in Aortic Ring from Normotensive Rats

Background: Ruthenium (Ru) tetraamines are being increasingly used as nitric oxide (NO) carriers. In this context, pharmacological studies have become highly relevant to better understand the mechanism of action involved. Objective: To evaluate the vascular response of the tetraamines trans-[RuII(NH3)4(Py)(NO)]3+, trans-[RuII(Cl)(NO) (cyclan)](PF6)2, and trans-[RuII(NH3)4(4-acPy)(NO)]3+. Methods: Aortic rings were contracted with noradrenaline (10−6 M). After voltage stabilization, a single concentration (10−6 M) of the compounds was added to the assay medium. The responses were recorded during 120 min. Vascular integrity was assessed functionally using acetylcholine at 10−6 M and sodium nitroprusside at 10−6 M as well as by histological examination. Results: Histological analysis confirmed the presence or absence of endothelial cells in those tissues. All tetraamine complexes altered the contractile response induced by norepinephrine, resulting in increased tone followed by relaxation. In rings with endothelium, the inhibition of endothelial NO caused a reduction of the contractile effect caused by pyridine NO. No significant responses were observed in rings with endothelium after treatment with cyclan NO. In contrast, in rings without endothelium, the inhibition of guanylate cyclase significantly reduced the contractile response caused by the pyridine NO and cyclan NO complexes, and both complexes caused a relaxing effect. Conclusion: The results indicate that the vascular effect of the evaluated complexes involved a decrease in the vascular tone induced by norepinephrine (10−6 M) at the end of the incubation period in aortic rings with and without endothelium, indicating the slow release of NO from these complexes and suggesting that the ligands promoted chemical stability to the molecule. Moreover, we demonstrated that the association of Ru with NO is more stable when the ligands pyridine and cyclan are used in the formulation of the compound.

Hypotensive Response Magnitude and Duration in Hypertensives: Continuous and Interval Exercise

Background: Although exercise training is known to promote post-exercise hypotension, there is currently no consistent argument about the effects of manipulating its various components (intensity, duration, rest periods, types of exercise, training methods) on the magnitude and duration of hypotensive response. Objective: To compare the effect of continuous and interval exercises on hypotensive response magnitude and duration in hypertensive patients by using ambulatory blood pressure monitoring (ABPM). Methods: The sample consisted of 20 elderly hypertensives. Each participant underwent three ABPM sessions: one control ABPM, without exercise; one ABPM after continuous exercise; and one ABPM after interval exercise. Systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), heart rate (HR) and double product (DP) were monitored to check post-exercise hypotension and for comparison between each ABPM. Results: ABPM after continuous exercise and after interval exercise showed post-exercise hypotension and a significant reduction (p < 0.05) in SBP, DBP, MAP and DP for 20 hours as compared with control ABPM. Comparing ABPM after continuous and ABPM after interval exercise, a significant reduction (p < 0.05) in SBP, DBP, MAP and DP was observed in the latter. Conclusion: Continuous and interval exercise trainings promote post-exercise hypotension with reduction in SBP, DBP, MAP and DP in the 20 hours following exercise. Interval exercise training causes greater post-exercise hypotension and lower cardiovascular overload as compared with continuous exercise.

Effects of neural nitric oxide synthase gene inactivation on the neurodocrine stress response in mice

Neural nitric oxide synthase, neuroendocrine stress response, forced swimming, nNOS KO mice, hypothalamus, adrenal gland