published_or_final_version / abstract / Biological Sciences / Master / Master of Philosophy

published_or_final_version / Biological Sciences / Master / Master of Philosophy

published_or_final_version / Medicine / Master / Master of Philosophy

published_or_final_version / Anatomy / Master / Master of Philosophy

published_or_final_version / Biochemistry / Master / Master of Philosophy

published_or_final_version / Biochemistry / Master / Master of Philosophy

published_or_final_version / Anatomy / Doctoral / Doctor of Philosophy

Removal of the pineal gland from partially parathyroidectomized rats produces stereotyped violent seizures. Inasmuch as the neurotransmitter norepinephrine (NE) has been implicated in this experimental paradigm, the purpose of this study was to investigate the effect of specific alterations in catecholamine function on convulsions produced by pinealectomy (PinX). Additionally, the role of various pineal substances, sex differences and the caging paradigm in the convulsive response was studied. Male and female rats (grouped five per cage) were found to respond similarly to the convulsive stimulus of parathyroidectomy followed by PinX. Neither implants of melatonin nor ventricular injections of arginine vasotocin in isolated and grouped rats, respectively, produced consistent changes in convulsions from PinX. The method of caging the rats after PinX, however, dramatically influenced seizures. Isolated rats (one per cage) convulsed significantly later after PinX and did so less often than grouped (five per cage) controls. NE neurotransmission appears to play a strong role in influencing PinX-induced seizures. Augmenting NE function with desipramine suppressed seizures. Convulsions were enhanced by the (beta)-receptor antagonist timolol, while neonatal injections of the catecholamine neurotoxin 6-OHDA potentiated seizures so markedly that many rats died from just one convulsion. NE levels were significantly reduced in the telencephalons and increased in the brain stems of sham-pinealectomized rats which had also received neonatal 6-OHDA; telencephalic levels of DA were elevated by 6-OHDA. Both the proconvulsant effects of 6-OHDA and the alterations it produced in central catecholamine levels were prevented, for the most part, by pretreatment with DMI. Altering both NE and DA function with L-dihydroxyphenylalanine, (alpha)-methyl-p-tyrosine, FLA-63 or reserpine did not significantly affect PinX-induced seizures in isolated rats. NE appears to play a strong role in modulating PinX-induced seizures; however, a deficit in NE function per se does not seem to be the fundamental cause of the seizures since sham-pinealectomized rats having lowered NE and/or DA function did not convulse.

La resynchronisation biventriculaire est un traitement recommandé chez les patients avec dysfonction ventriculaire gauche (VG) sévère et QRS large. Si les résultats en termes de bénéfice clinique sont globalement positifs, toutes les études retrouvent un pourcentage non négligeable de patients non répondeurs à la thérapeutique. Dans ce travail, en couplant données expérimentales animales et études cliniques, nous avons 1) investigué l’impact de la resynchronisation biventriculaire chez des patients avec Tétralogie de Fallot opérée 2) évalué l’impact hémodynamique associé avec une stimulation VG multipoints et avec une stimulation VG endocardique 3) analysé l’intérêt de l’optimisation des paramètres de stimulation à l’effort. Nous avons mis en évidence que 1) la resynchronisation permet un bénéfice hémodynamique significatif sur un modèle animal de dysfonction ventriculaire droite et chez des patients avec Tétralogie de Fallot opérée 2) la stimulation multipoints et la stimulation endocardique VG permettent un bénéfice hémodynamique significatif sur des modèles animaux d’insuffisance cardiaque et chez des patients avec insuffisance cardiaque sévère 3) l’optimisation à l’effort des paramètres de programmation permet un bénéfice hémodynamique. Un capteur intégré dans la prothèse de stimulation pourrait permettre une optimisation automatique.L’ensemble de ces données permet d’espérer une optimisation de la réponse clinique et d’envisager de nouveaux champs d’application pour cette thérapeutique. / Cardiac resynchronization therapy is recommanded in patients with severe left ventricular (LV) dysfunction and wide QRS. Despite positive clinical results, a significant proportion of implanted patients do not demonstrate positive response to the therapy. Coupling experimental data and clinical studies, we have 1) investigated the impact of cardiac resynchronization in patients with repaired Tetralogy of Fallot 2) assessed the hemodynamic impact associated with multisite LV pacing and endocardial LV pacing 3) analyzed the impact of an exercise-optimization of the programmed parameters.We have demonstrated that 1) biventricular pacing is associated with a significant hemodynamic improvement in an animal model of right ventricular dysfunction and in patients with repaired Tetralogy of Fallot 2) multisite LV pacing and endocadial LV pacing are associated with significant hemodynamic improvement in animal models and in humans with severe heart failure 3) optimization during exercise of AV and VV delays induce acute hemodynamic improvement and could be automatically performed by an integrated hemodynimc sensor. Our data suggest a posible improvement in clinical response after cardiac resynchronization and a posible extension of the indications.

The general aim of the thesis was to deduce whether selected naturally occurring flavonoids (genistein, epicatechin gallate (EC), epigallocatechin gallate (EGCG), baicalin) attenuate various secondary insults that may cause death of ganglion cells in primary open angle glaucoma (POAG). An ischemic insult to the rat retina significantly causes the inner retina to degenerate indexed by changes of various antigens, proteins and mRNAs located to amacrine and ganglion cells. These changes are blunted in animals treated with genistein as has been shown for ECGC. Studies conducted on cells (RGC-5 cells) in culture showed that hydrogen peroxide, L-buthionine sulfoximine (BSO)/glutamate and serum deprivation (mimicking oxidative stress), rotenone, sodium azide (affecting mitochondria function in specific ways) and light (where the mitochondria are generally affected) all generated reactive oxygen species and caused death of RGC-5 cells. EGCG was able to attenuate cell death caused by hydrogen peroxide, sodium azide and rotenone. Only EC was able to attenuate BSO/glutamate-induced cell death, in addition to cell death caused by hydrogen peroxide and rotenone. Genistein had no positive effect on cell death in experiments carried out on RGC-5 cells. Exposure of RGC-5 cells to flavonoids showed that EC and EGCG increased the mRNA expression of endogenous antioxidants such as HO-l (heme oxygenase 1) and Nrf-2 (nuclear erythroid factor-z-related factor 2). Light insult, rotenone and sodium azide activate the p38 (protein kinase 38) pathway, while only light and rotenone activate the JNK (c-Jun amino-terminal kinase) pathway. Serum deprivation affects mitochondrial apoptotic proteins causing an increase in the ratio of Bax/Bcl2 (Bax: Bcl-2-associated X protein; Bcl-2: B-cell lymphoma 2). An insult of light to RGC-5 cells, unlike that induced by sodium azide, is inhibited by necrostatin-I and causes an activation of AlF (apoptosis-inducing factor) with alpha-fodrin being unaffected. These studies suggest that ganglion cell death caused by insults as may occur in POAG involves various cellular signaling pathways. The selected flavonoids have diverse actions in increasing cellular defense mechanisms, and in negating the effects of ischemia and specific types of oxidative stress. The results argue for the possible use of flavonoids in the treatment of POAG to slow down ganglion cell death.