No description available.

Cardiac disease is one of the leading causes of death in the world, with an increase in cardiac arrhythmias in recent years. In addition, myocardial ischemia, which arises from the lack of blood in the cardiac tissue, can lead to cardiac arrhythmias and even sudden cardiac death. Cardiac arrhythmias, such as atrial fibrillation, are characterised by abnormal wave excitation and repolarization patterns in the myocardial tissue. These abnormal patterns are usually diagnosed through non-invasive electrical measurements on the surface of the body, i.e., the electrocardiogram (ECG). However, the most common lead configuration of the ECG, the 12-lead ECG, has its limitations in providing sufficient information to identify and locate the origin of cardiac arrhythmias. Therefore, there is an increasing need to develop novel methods to diagnose and find the origin of arrhythmic excitation, which will increase the efficacy of the treatment and diagnosis of cardiac arrhythmias. The objective of this research was to develop a family of multi-scale computational models of the human heart and thorax to simulate and investigate the effect of arrhythmic electrical activity in the heart on the electric and magnetic activities on the surface of the body. Based on these simulations, new theoretical algorithms were developed to non-invasively diagnose the origins of cardiac arrhythmias, such as the location of ectopic activities in the atria or ischemic regions within the ventricles, which are challenging to the clinician. These non-invasive diagnose methods were based on the implementation of multi-lead ECG systems, magnetocardiograms (MCGs) and electrocardiographic imaging.

No description available.

Background: Extensive research has characterized the embryonic development of a four-chambered heart in mammals. After birth, mammalian cardiomyocytes undergo a transition characterized by a final cell cycle with nuclear division (karyokinesis) in the absence of cytoplasmic division (cytokinesis), generating mature binucleated cardiomyocytes. Downregulation of pro-proliferative signaling and epigenetic changes permanently ‘lock’ cardiomyocytes out of the cell cycle, and nearly all subsequent growth is accomplished via cellular hypertrophy. Before this transition, cardiomyocytes exhibit robust proliferative potential, but afterward are unable to divide. Rationale & Hypothesis: Recent evidence suggests that non-coding RNAs influence early neonatal cardiac development and hypertrophy. We hypothesize that transient expression of regulatory miRNAs may impact the neonatal heart’s transition from proliferation to hypertrophy. Results: Cardiac mRNA and miRNA were systematically analyzed using microarrays to identify targets that were transiently and significantly changing after birth. Through our analysis we identified three primary ontogenies significantly changing: metabolism, extracellular matrix remodeling, and cell cycle regulation. Global analysis of micro-RNA expression patterns during perinatal heart development identified miR-205 as a novel candidate for modulating cardiomyocyte maturation. We observed miR-205 expression undergoing a 20-fold increase from 1-day postpartum (1D) to 5D, returning to prenatal levels by 10D. It is expressed in cardiomyocytes of the epicardium, the primary location of fetal cardiomyocyte proliferation. MiR-205 targets two important cell cycle regulators: Pten phosphatase of the PI3K/AKT pathway, and Yap1 in the Hippo pathway. Both pathways have proven to be essential for proper heart development. Previous research showed that germline deletion of miR-205 results in death at 5D. To define its role in the heart, we generated an αMHC-Cre postnatal miR-205 cardiac-specific deletion mouse model. Systematic characterization of miR-205-/- hearts confirmed miR-205’s interaction with Pten and Yap1 by western blot and immunohistochemistry. Postnatal miR-205-/- hearts exhibit Hippo pathway dysregulation, increased cardiomyocyte number, more actively cycling cardiomyocytes beyond 7D, and no difference in binucleation. We also generated a DOX-inducible cardiac-specific miR-205 over-expression mouse model. Perinatal miR-205OE hearts expedited the transitional period, with more cardiomyocytes present at 5D and no difference at 14D. These hearts show increased Hippo signaling immediately after birth, suggesting compensatory mechanisms to ensure sufficient cardiomyocyte number. Conclusions: Our data strongly supports miR-205 as a regulator of cardiomyocyte maturation in the neonatal heart, by promoting the neonatal cardiomyocyte transition from hyperplastic to hypertrophic growth. In turn, miR-205’s antiproliferative properties originate in part from suppressing the expression of Pten and Yap1.

Abstract not available

Hypertension is one of many side effects of oral contraceptive use in a small percentage of women. Although the underlying pathology has yet to be fully resolved, alterations in the renin-angiotensin-aldosterone axis, sympathetic nervous system/ renal and cardiac function have been implicated. In the thesis to be presented, the possible involvement of alterations in renal and myocardial adrenoceptor characteristics in the pathogenesis of steroid contraceptive-induced hypertension in rats was examined by radioligand binding techniques. In Chapter 2, a rat model of OC hypertension is described. Chronic low-dose administration of ethynyloestradiol (EE2), levonorgestrel (NG) or a combination of both steroids (EE2/NG) to female Sprague-Dawley rats was shown to significantly increase systolic blood pressure (SBP). Renal and cardiac hypertrophy developed in association with EE2-, EE2/NG- but not NG-induced hypertension. Moreover, whereas administration of NG alone attenuated body weight gain, combined EE2/NG administration increased body weight gain from the second week of treatment onwards. Based on the above observations, it is proposed that EE2 and NG induce hypertension in rats via different mechanisms. Although SBP was elevated to a similar maximum in all steroid-treated groups (+ 20 mmHg compared to controls), only with EE2 administration did SBP remain elevated for the duration of the 17 week treatment regimen. NG may therefore have a protective effect on blood pressure with long-term combined steroid contraceptive treatment. In Chapter 4, renal adrenoceptors were characterized using radioactively labelled adrenocephor antagonists. Under appropriate conditions, binding of [3H]-prazosin and [3H]-rauwolscine to membrane preparations of whole rat kidney displayed the kinetics, saturability and specificity of α1- and α2 -adrenoceptors respectively, which were present in a ratio 3:1. In contrast, [3H]-dihydroergocryptine ([3H]-DHE) apparently bound to both α1 and α2-adrenoceptors. Binding sites identified by [125I] –iodocyanopindolol (ICYP) had the recognition characteristics of β-adrenoceptors. In drug competition studies using the subtype-selective antagonists practolol (β1) and ICI 118,551 (β2)/ the ratio of β1- to β2 -adrenoceptors was found to be approximately 2:1. Subsequently, renal adrenoceptors were investigated at various stages during the development of hypertension with the different steroid contraceptive treatments (Chapters 5 and 6). Preliminary binding studies with [3H]-DHE and [3H]-prazosin suggested that the number of renal α2 - but not α1-adrenoceptors was reduced in rats with established EE2-induced hypertension (17 weeks treatment). This was subsequently confirmed using [3H]-rauwolscine, which in addition showed that the reduction in renal α2 -adrenoceptor number occurred during the developmental stage of EE2/NG~induced hypertension (6 weeks treatment) and established EE2-induced hypertension (12 weeks treatment). NG induced hypertension was unassociated with changes in renal α1- and α2-adrenoceptor characteristics. Renal β-adrenoceptor affinity was reduced in established EE2-, but not NG- or EE2/NG- induced hypertension. Moreover, the β-adrenoceptor agonist (-)-isoprenaline bound to renal β-adrenoceptors with reduced affinity following EE2 administration. Several endogenous and synthetic steroids were found to be ineffective inhibitors of [3H] –prazosin, [3H] –rauwolscine and ICYP binding excluding a direct interaction of these steroids with renal α1-, α2- and β -adrenoceptors. In Chapter 7, myocardial adrenoceptors were characterized and investigated in steroid-treated rats. In membrane preparations of whole myocardium, [3H]-prazosin binding was characteristically to α1- adrenoceptors, whereas there was a notable absence of [3H]-rauwolscine binding. Using ICYP, β-adrenoceptors were also detected, the ratio of β1- to β2~adrenoceptors being 3:1. Steroid contraceptive-induced hypertension was not associated with myocardial α1-adrenoceptor changes. Similarly, myocardial β-adrenoceptors were unchanged in established EE2-, NG- and EE2/NG-induced hypertension (12 weeks treatment). The affinity of (-)-isoprenaline for myocardial β-adrenoceptors was unaffected by EE2 aditiinistration. These studies suggest that established EE2- but not NG-induced hypertension in rats is associated with selective alterations in renal α2- and (β-adrenoceptors. These adrenoceptor changes may help to maintain elevated blood pressure by affecting the control of renal function by the sympathetic nervous system, catecholamines and several hormones which affect renin release and the transport of fluid and electrolytes in the nephron.

8-Cyclopentyl-3-(3-(4-fluorosulfonylbenzoyl)oxy)propyl-propylxanthine (44, FSCPX) has been reported to exhibit potent and selective irreversible antagonism of the A1 adenosine receptor when using in vitro biological preparations. However, FSCPX (44) suffers from cleavage of the ester linkage separating the reactive 4-(fluorosulfonyl)phenyl moiety from the xanthine pharmacophore when used in in vivo biological preparations or preparations containing significant enzyme activity, presumably by esterases. Cleavage of the ester linkage renders FSCPX (44) inactive in terms of irreversible receptor binding. In order to obtain an irreversible A1 adenosine receptor antagonist with improved stability, and to further elucidate the effects of linker structure on pharmacological characteristics, several FSCPX (44) analogues incorporating the chemoreactive 4-(fluorosulfonyl)phenyl moiety were targeted, where the labile ester linkage has been replaced by more stable functionalites. In particular, ether, alkyl, amide and ketone linkers were targeted, where the length of the alkyl chain was varied from between one to five atoms. Synthesis of the target compounds was achieved via direct attachment of the N-3 substituent to the xanthine. These compounds were then tested for their biological activity at the A1 adenosine receptor via their ability to irreversibly antagonise the binding of [3H]-8-cyclopentyl-1,3-dipropylxanthine ([3H]DPCPX, ( 9) to the A1 adenosine receptor of DDT1 MF-2 cells. For comparison, the xanthines were also tested for their ability to inhibit the binding of [3H]-4-(2-[7-amino-2-{furyl} {1,2,4}- triazolo{2,3-a} {1,3,5}triazin-5-ylamino-ethyl)]phenol ([3H]ZM241385, 36) to the A2A adenosine receptor of PC-12 cells. The results suggest that the length and chemical composition of the linker separating the reactive 4-(fluorosulfonyl)phenyl moiety from the xanthine ring contribute to the potency and efficacy of the irreversible A1 adenosine receptor ligands. Like FSCPX (44, IC50 A1 = 11.8 nM), all derivatives possessed IC50 values in the low nM range under in vitro conditions. Compounds 94 (IC50 A1 = 165 nM), 95 (IC50 A1 = 112 nM) and 96 (IC50 A1 = 101 nM) possessing one, three and five methylene spacers within the linkage respectively, exhibited potent and selective binding to the A1 adenosine receptor versus the A2A adenosine receptor. Compound 94 did not exhibit any irreversible binding at A1 adenosine receptors, while 95 and 96 exhibit only weak irreversible binding at A1 adenosine receptors. Those compounds containing a benzylic carbonyl separating the 4-(fluorosulfonyl)phenyl moiety from the xanthine ring in the form of an amide (119, IC50 A1 = 24.9 nM, and 120, IC50 A1 = 21 nM) or ketone (151, IC50 A1 = 14 nM) proved to be the most potent, with compound 120 exhibiting the highest selectivity of 132-fold for the A receptor over the A2A receptor. compounds 119, 120 and 151 also strongly inhibited the binding of [3H]DPCPX irreversibly (82%, 83% and 78% loss of [3H]DPCPX binding at 100 nM respectively). compounds 120 and 151 are currently being evaluated for use in in vivo studies. Structure-activity studies suggest that altering the 8-cycloalkyl group of A1 selective xanthines for a 3-substituted or 2,3-disubstituted styryl, combined with N-7 methyl substitution will produce a compound with high affinity and selectivity for the A2A adenosine receptor over the A1 adenosine receptor. Compound 167 (IC50 A2A = 264 nM) possessing 8-(m-chloro)styryl substitution and the reactive 4-(fluorosulfonyl)phenyl moiety separated from the xanthine ring via an amide linker in the 3-position (as for 119 and 120), exhibited relatively potent binding to the A2A adenosine receptor of PC-12 cells, with a 16-fold selectivity for that receptor over the A1 adenosine receptor. However, compound 167 exhibited only very weak irreversible binding at A2A adenosine receptors. Overall, at this stage of biological testing, compound 120 appears to possess the most advantageous characteristics as an irreversible antagonist for the A1 adenosine receptor. This can be attributed to its high selectivity for the A1 adenosine receptor as compared to the A2A adenosine receptor. It also has relatively high potency for the A1 adenosine receptor, a concentration-dependent and selective inactivation of A1 adenosine receptors, and unbound ligand is easily removed (washed out) from biological membranes. These characteristics mean compound 151 has the potential to be a useful tool for the further study of the structure and function of the A1 adenosine receptor.

<p>Methods for measuring deformation and motion of the human heart in-vivo are crucial in the assessment of cardiac function. Applications ranging from basic physiological research, through early detection of disease to follow-up studies, all benefit from improved methods of measuring the dynamics of the heart. This thesis presents new methods for acquisition, reconstruction and visualization of cardiac motion and deformation, based on magnetic resonance imaging.</p><p>Local heart wall deformation can be quantified in a strain rate tensor field. This tensor field describes the local deformation excluding rigid body translation and rotation. The drawback of studying this tensor-valued quantity, as opposed to a velocity vector field, is the high dimensionality of the tensor. The problem of visualizing the tensor field is approached by combining a local visualization that displays all degrees of freedom for a single tensor with an overview visualization using a scalar field representation of the complete tensor field. The scalar field is obtained by iterated adaptive filtering of a noise field.</p><p>Several methods for synchronizing the magnetic resonance imaging acquisition to the heart beat have previously been used to resolve individual heart phases from multiple cardiac cycles. In the present work, one of these techniques is extended to resolve two temporal dimensions simultaneously, the cardiac cycle and the respiratory cycle. This is combined with volumetric imaging to produce a five-dimensional data set. Furthermore, the acquisition order is optimized in order to reduce eddy current artifacts.</p><p>The five-dimensional acquisition either requires very long scan times or can only provide low spatiotemporal resolution. A method that exploits the variation in temporal bandwidth over the imaging volume, k-t BLAST, is described and extended to two simultaneous temporal dimensions. The new method, k-t2 BLAST, allows simultaneous reduction of scan time and improvement of spatial resolution.</p> / Report code: LIU-TEK-LIC-2006:43

Background: Decreased right ventricular function after coronary artery bypass graft surgery is a well-known phenomenon. The use of an artificial circuit and of cardioplegia during cardiopulmonary bypass (conventional coronary artery bypass graft surgery) has been evoked as possible reason for this dysfunction. With the availability of cardiac stabilisers (Octopus or Starfish®), bypass surgery on a beating heart is now possible without the use of cardioplegia and cardiopulmonary bypass (off-pump coronary artery bypass graft surgery). Avoiding the potential damaging effects of cardiopulmonary bypass and keeping a nearly continuous perfusion in the coronary arteries of the beating heart have been postulated to offer a better protection of the myocardium with a better outcome and to reduce the perioperative morbidity. Aims: If right ventricular dysfunction is known to occur after conventional coronary artery bypass graft surgery, nothing is known about right ventricular function after off-pump coronary artery bypass graft surgery. Therefore we performed two prospective, randomised, controlled trials to assess and compare the short-term and medium-term effects of conventional and off-pump coronary artery bypass graft surgery on the global and regional right ventricular function. We used transoesophageal and transthoracic echocardiography as investigating tools of right ventricular systolic and diastolic function. Materials and Methods: Fifty consecutive patients scheduled for elective coronary bypass graft surgery and for whom the surgeon regarded off-pump and on-pump techniques as equally suitable were randomised. We performed a transthoracic echocardiography the day before surgery and 3 months after surgery, and a transoesophageal echocardiography just before opening and just after closure of the sternum. Results: Just after closure of the sternum, we could not observe a better protection of the right ventricular function by the off-pump surgery. Cardiac index and right ventricular ejection fraction were equally preserved in the 2 groups. Systolic and diastolic myocardial function (investigated by Tissue Doppler imaging) was impaired only in the off-pump group, but the intergroup difference was not statistically significant. Three months after surgery, there were no differences between the 2 groups: global right ventricular systolic function was equally preserved and right ventricular systolic and diastolic myocardial function equally impaired. Conclusions: These 2 studies do not allow for concluding that off-pump coronary artery bypass graft surgery would better preserve right ventricular systolic and diastolic function than conventional coronary artery bypass graft surgery, just after surgery or 3 months later. In any event, our studies provided the opportunity to acquire experience in systematically evaluating the right ventricle using transoesophageal echocardiography. / Introduction: La dysfonction ventriculaire droite après chirurgie coronaire est un phénomène bien connu. L'utilisation d'un circuit artificiel ainsi que d'une solution de cardioplégie pendant la circulation extra-corporelle (CEC) (chirurgie coronaire classique) a été évoquée comme origine possible de cette dysfonction. Une chirurgie sur cœur battant (chirurgie coronaire sans CEC) est actuellement possible grâce à l'utilisation de stabilisateurs cardiaques (Octopus ou Starfish®). Ce type de chirurgie sans CEC permet d'éviter l'utilisation de la CEC et de solutions de cardioplégie et d'en supprimer ainsi les effets délétères. Elle permet également de maintenir une perfusion quasi continue des artères coronaires. Ces différents éléments ont été évoqués comme offrant une meilleure protection myocardique ainsi qu'un meilleur devenir des patients, et comme pouvant réduire la morbidité périopératoire. Buts : La survenue d'une dysfonction ventriculaire droite après chirurgie coronaire classique est un phénomène bien connu, par contre nous savons peu de chose sur la fonction ventriculaire droite après chirurgie coronaire sans CEC. Pour cette raison, nous avons entrepris deux études prospectives, randomisées et contrôlées afin d'étudier et de comparer les effets à court et moyen terme de la chirurgie coronaire classique et sans CEC. Nous avons utilisé l'échographie cardiaque transthoracique et transœsophagienne comme outil de mesure de la fonction systolique et diastolique du ventricule droit. Matériel et Méthode : Nous avons randomisé cinquante patients consécutifs, programmés pour une chirurgie coronaire élective et pour laquelle le chirurgien estimait qu'une chirurgie sans CEC était aussi appropriée qu'une chirurgie avec CEC. Une échographie cardiaque transthoracique était réalisée la veille de l'intervention ainsi que 3 mois plus tard, une échographie transoesophagienne était réalisée juste avant l'ouverture et après la fermeture du sternum. Résultats : Juste après la fermeture du sternum, la fonction ventriculaire droite dans le groupe sans CEC n'était pas mieux protégée que dans le groupe avec CEC. L'index cardiaque et la fraction d'éjection ventriculaire droite étaient préservés de manière équivalente dans les 2 groupes. La fonction régionale systolique et diastolique du ventricule droit (étudiée par Doppler tissulaire) n'était altérée de manière significative que dans le groupe sans CEC, mais la différence intergroup n'était pas statistiquement significative. A 3 mois également, les 2 groupes ne différaient pas : la fonction systolique globale du ventricule droit était préservée de manière équivalente ; la fonction régionale systolique et diastolique du ventricule droit était altérée de manière équivalente. Conclusions : Ces 2 études ne nous permettent pas de conclure à une meilleure protection de la fonction systolique et diastolique ventriculaire droite par la chirurgie coronaire sans CEC, que ce soit juste en fin de chirurgie ou 3 mois plus tard. D'autre part, ces études nous ont permis d'acquérir de l'expérience dans l'analyse systématique du ventricule droit par échographie cardiaque transoesophagienne.

During embryonic development, multipotent cardiovascular progenitor cells (MCPs) are specified from early mesoderm. Although the core cardiac transcriptional machinery acting during cardiac cell differentiation is relatively well known, the molecular mechanism acting upstream of these cardiac transcriptional factors, and promoting cardiac progenitor specification from early mesoderm remains poorly understood. We used embryonic stem cell (ESC) differentiation as a model to dissect the molecular mechanisms implicated in cardiovascular progenitor specification. Using ESCs, in which gene expression can be temporally regulated, we showed that transient expression of Mesp1 dramatically accelerates and enhances multipotent cardiovascular progenitor specification through an intrinsic and cellular autonomous mechanism. Using genome wide transcriptional analysis, we found that Mesp1 rapidly activates and represses a discrete set of genes. Using chromatin immunoprecipitation, we showed that Mesp1 directly binds to regulatory DNA sequences located in the promoter of many key genes belonging to the core cardiac transcriptional machinery, resulting in their rapid upregulation. Mesp1 also directly and strongly represses the expression of key genes regulating other early mesoderm and endoderm cell fates. Using engineered ESC expressing the green fluorescent protein under the control of the Mesp1 promoter, we isolated Mesp1 expressing cells in differentiating ESCs allowing characterization of the cellular and molecular mechanisms underlying cardiovascular specification. Our results demonstrate that Mesp1 acts as a key regulatory switch during cardiovascular specification, residing at the top of the hierarchy of the gene network responsible for cardiovascular cell fate determination. Moreover our results place Mesp1 upstream of the specification of both first and second heart fields and provide novel and important insights into the molecular mechanisms underlying the earliest step of cardiovascular specification. We identified cell surface markers expressed allowing the isolation of early cardiovascular progenitors and provide potentially novel methods for dramatically increasing the number of cardiovascular cells for cellular therapy in humans.