Pharmacokinetic/pharmacodynamic relationship of pulmonary administration methods for milrinone : a translational approach

Suite à l'interruption de la circulation extracorporelle (CEC) chez les patients qui subissent une chirurgie cardiaque, le rétablissement de la circulation sanguine au niveau du poumon cause souvent une hypertension pulmonaire qui, à son tour, peut engendrer des complications au cœur droit. Pour traiter cette hypertension, la milrinone, un inhibiteur de la phosphodiestérase 3 (PDEIII) est souvent donnée par voie intraveineuse. Par contre, cette administration cause également une hypotension systémique qui peut nécessiter l’administration d'agents vasoactifs. Une administration de la milrinone par nébulisation a donc été adoptée, ciblant ainsi directement les poumons [1]. En cas d'urgence, la milrinone peut être injectée ou atomisée directement au niveau de la trachée. Lors de chirurgies cardiaques nécessitant une CEC, un protocole expérimental s'avère souvent difficile à respecter chez le patient. En effet, les contraintes temporelles, les nombreux médicaments co-administrés et le stress physique causé par la chirurgie sont autant de facteurs qui peuvent biaiser une étude dont le but est caractériser l’effet pharmacologique de la milrinone. Il s'est donc avéré important, en premier lieu, de quantifier les effets des artéfacts reliés aux manipulations chirurgicales sur notre biomarqueur, soit le ratio de la pression systémique moyenne sur la pression pulmonaire moyenne. Chez le porc, nous avons observé des changements de 30-50% au niveau des pressions systémiques moyennes, soit le double des changements au niveau des pressions pulmonaires pour le même artéfact. La durée moyenne de tous les effets, des artéfacts, (au niveau systémique et pulmonaire conjointement) fut de 4.5±2.5 min (n=152). Dû au biais potentiel apporté par ces artéfacts, l’utilisation d’un modèle animal dépourvu de manipulations chirurgicales s’est avérée être pleinement justifiée. En second lieu, la voie pulmonaire étant une voie d'administration non homologuée pour la milrinone, le site optimal de déposition pulmonaire permettant un effet local maximal n’a pas été identifié. La milrinone cible la PDEIII au niveau des muscles lisses vasculaires. Ainsi, l’effet anti-hypertensif local de la milrinone se situe au niveau des artères musculaires côtoyant l’arbre trachéobronchique. Nos études se concentrent sur les nébulisations par simple jet (SJ), par mèche vibrante (VM) ainsi que sur les administrations d’urgence, soit l’instillation (ITI) et l’atomisation (ITA) intratrachéale. De ce fait, nous avons conduit des études in vitro pour caractériser la taille des particules générée par les deux formes de nébulisation, qui est un facteur influençant la déposition pulmonaire. Nous avons aussi tenté, grâce à des études in et ex vivo, de démontrer une différence d’efficacité entre les deux méthodes de nébulisation. Les études in vitro ont permis, de quantifier la dose se rendant au poumon, soit la dose inhalée, et ont démontré une différence au niveau de la distribution de la taille des particules des gouttelettes générées par les deux nébulisations. Les résultats indiquent une taille de particules plus fine, visant préférentiellement la zone alvéolaire pour la nébulisation par mèche vibrante. Ils indiquent aussi une plus grande dose inhalée pour la nébulisation par mèche vibrante mais de façon surprenante, en tenant compte de la dose inhalée par le patient, une meilleure relaxation pulmonaire par simple jet. En plus d’une meilleure oxygénation suite à l’administration de la milrinone, nos études démontrent une sensibilité de 20 fois plus grande, tel que démontrée par le IC50 de la relaxation à l’acétylcholine, et un effet de relaxation près de 25% supérieur chez la nébulisation par simple jet En troisième lieu, la relation pharmacocinétique (PK) et pharmacodynamie (PD) de la milrinone par nébulisation n'a pas été caractérisée en absence d’artéfacts chirurgicaux et encore moins pour les voies d’urgence. Suite au développement d’un modèle d’hypertension pulmonaire sans manipulations chirurgicales, nous avons déterminé une relation concentration/effet (PK/PD) pour les méthodes d’administration pulmonaire et comparé leurs efficacités après correction pour la dose réellement inhalée, déterminée par collecte urinaire. Cette dernière étude démontre un EC50 de l’impact de la milrinone sur le ratio des pressions artérielles systémiques moyennes sur les pressions artérielles pulmonaires plus faible suite à l’administration de la milrinone par simple jet ainsi qu’une augmentation de ce ratio de près 40%. Les résultats obtenus au cours de ces études suggèrent l’administration par simple jet semble plus efficace. Par conséquent, un meilleur jugement clinique pourra être appliqué lors de la méthode d’administration ainsi que de la dose utilisée de la milrinone.Following weaning from cardiopulmonary bypass (CPB), the cardiac surgery patients undergoing a procedure are often faced with the pulmonary reperfusion syndrome which may lead to pulmonary hypertension condition which, in turn, is likely to precipitate right ventricular dysfunction. Milrinone, a phosphodiesterase type 3 inhibitor (PDEIII) is used, among others, to facilitate CPB weaning in patients by reducing the severity of pulmonary hypertension through vasodilation. Milrinone is typically given intravenously. However, this mode of administration is frequently associated with systemic hypotension which may require administration of vasoactive agents. Administration of milrinone throughnebulization has therefore been developed, targeting the lungs directly and avoiding systemic hypotension. In case of emergency, milrinone may even be injected directly into the trachea or even possibly atomized intratrachealy. In this thesis we will describe an experimental protocol where the administration of milrinone through inhalation was studied. Unfortunately, experimental protocols during surgeries requiring CPB are often challenging due to time and procedural constraints. Indeed, the signal-to-noise ratio makes it difficult to isolate the effect of a specific intervention. The effect of the other administered medications and the physical stress of surgery may result in potentially incomplete or biased pharmacological studies. This aspect is the basis of the first portion of the thesis. The identification and and the quantification of the surgical manipulation artifacts on our main biomarker, the mean systemic pressure over the mean pulmonary pressure, was our first objective. In our first study, following our experimentation, we witnessed a 30- 50% changes on systemic pressures and these changes were twice those observed on pulmonary pressures for the same artefact. Pooled durations of all effects on both mAP and mPAP averaged 4.5±2.5 min (n=152). These changes fully justified the creation of a new model due to the potential biais of the artefacts. Secondly, as nebulization is a novel way to administer milrinone, the pharmacokinetic-pharmacodynamics (PK-PD) relationship has not been completely characterized. Even less information is available for the emergency routes. Milrinone targets PDEIII in vascular smooth muscle, thus the local antihypertensive effect of milrinone is most likely to occur at the level of muscular arteries alongside the tracheobronchial tree. We studied four methods of administration: simple jet (SJ), vibrating mesh (VM) nebulization, intratracheal instillation (ITI) and atomization (ITA). In addition, the optimal pulmonary deposition and absorption site for maximum local effect remains as of yet unknown. We have strived to identify the factors influencing the lung deposition of milrinone through nebulization and part of the work discussed in this thesis is aimed at characterizing the amount of milrinone deposited on the various sections of the tracheobronchial tree by the mentioned methods. Through in and ex vivo studies we have investigated the differences in efficacy of the two nebulization methods. We have found each method to generate a significantly different deposition pattern and as such, influencing lung deposition. Our results show that VM targets the lower airways and alveoli, whereas SJ targets preferably the higher and middle airways. Our study also shows a higher inhaled dose for VM but surprisingly a better dose-related efficacy for SJ. Indeed, in addition to a better oxygenation, our study shows, as demonstrated by the acetylcholine-induced relaxation IC50, a 20 times higher sensitivity and a 25% higher maximum relaxation following simple jet nebulization. Subsequently, following the development of a pulmonary hypertension model devoid of surgical manipulations and, by association, artefacts, we have determined a urinary-dose-corrected PK/PD relationship for the nebulization and emergency methods of milrinone administration. We explored the methods of intratracheal bolus and atomization and compared their efficacy to those of nebulization. This study shows a lower EC50 of milrinone impact and an almost 40% increase of the mean systemic arterial pressure over mean pulmonary arterial pressures ratio following simple jet adminstration. The information obtained by these studies offers the potential to aid clinicians in making a more informed judgment in the use of milrinone

Revisiting the Sequence Method for Baroreflex Analysis

The sequence method is an important approach to assess the baroreflex function, mainly because it is based on the spontaneous fluctuations of beat-by-beat arterial pressure (for example, systolic arterial pressure or SAP) and pulse interval (PI). However, some studies revealed that the baroreflex effectiveness index (BEI), calculated through the sequence method, shows an intriguing oscillatory pattern as function of the delay between SAP and PI. It has been hypothesized that this pattern is related to the respiratory influence on SAP and/or PI variability, limiting the SAP ramps to 3 or 4 beats of length. In this study, this hypothesis was tested by assessing the sequence method using raw (original) and filtered series. Results were contrasted to the well-established transfer function, estimated between SAP and PI. Continuous arterial pressure recordings were obtained from healthy rats (N = 61) and beat-by-beat series of SAP and PI were generated. Low-pass (LP) and high-pass (HP) filtered series of SAP and PI were created by filtering the original series with a cutoff frequency of 0.8 Hz. Original series were analyzed by either the sequence method or cross-spectral analysis (transfer function) at low- (LF) and high- (HF) frequency bands, while filtered series were evaluated only by the sequence method. Baroreflex sensitivity (BRS) and BEI of original series, calculated by sequence method, was highly (85–90%) determined by HP series, with no significant association between original and LP series. A high correlation (>0.7) was found between the BRS estimated from original series (sequence method) and HF band (transfer function), as well as for LP series (sequence method) and LF band (transfer function). These findings confirmed the hypothesis that the sequence method quantifies only the high-frequency components of the baroreflex, neglecting the low-frequency influences, such as the Mayer waves. Therefore, we propose using both the original and LP filtered time series for a broader assessment of the baroreflex function using the sequence method

The new old normal : reassessing perioperative oxygen consumption and haemodynamics in the elderly

Perioperative haemodynamic instability and disturbances of global oxygen transport are associated with complications and organ injury after surgery. The continuously growing population of elderly in surgical care are at higher risk due to age-related cardiovascular alterations and increased prevalence of comorbidities. Optimised and tailored haemodynamic interventions may improve outcomes, but goals to aim for and responses to expect are not adjusted for elderly. Hypotension and changes in oxygen consumption (VO2) induced by anaesthesia are potentially very relevant in the elderly and reassessment is needed in modern perioperative care with current methodologies. In this thesis, cardiac output and haemodynamic changes related to hypotension after spinal anaesthesia (SPA) are outlined in the first study. VO2 after general anaesthesia (GA) and surgery is investigated in three studies with different approaches; by meta-analysis, prospectively during major surgery and by method comparison. Study I (prospective observational): 20 ASA II-IV patients (mean age 72 years) were monitored with LiDCO™plus prior to and 45 minutes after injection of SPA. Stroke volume and cardiac index, and consequently oxygen delivery index, decreased before the intrathecal injection and this decrease progressed after SPA in those who developed hypotension. In contrast, the non-hypotensive demonstrated an initial increase in cardiac index after SPA. Logistic regression analyses demonstrated that pre-anaesthetic changes of cardiac index could predict post-spinal hypotension (OR 0.79, 95% CI: 0.60, 0.91) with high discriminative ability (AUC 0.91). Study II (systematic review and meta-analysis): Cochrane Library, MEDLINE and EMBASE databases were searched for studies with measurements of VO2 before and after induction of GA. 24 studies with 453 patients were identified, published 1969-2000. Cochrane and NIH quality assessment tools revealed general high risk of bias in the majority of studies. However, measurements and interventions were described in great detail. A random-effects meta-analysis estimated the reduction of VO2 to -33 (95% CI: -38, -28) ml min-1 m-2 during GA but with uncertainty of the estimate due to very low quality as indicated in a GRADE evidence profile. A sample size calculation for study III was performed based on this data. Study III (prospective observational): VO2 was measured by indirect calorimetry (QuarkRMR), before, during and after major upper abdominal surgery in 20 ASA II-IV patients (mean age 73 years). VO2 decreased by a mean of -46 (95% CI: -55, -38) ml min-1 m-2 after induction of GA and increased during surgery. Simultaneous calculations of oxygen delivery (DO2) and estimated oxygen extraction ratio (O2ER) from LiDCO™plus monitoring and arterial-central venous blood gas content showed low intraoperative levels of extraction and delivery. Mixed effect models of relative changes of intraoperative VO2 compared to DO2 and estimated O2ER indicated that these parameters changed in parallel. Study IV (method comparison): Estimations of VO2 by LiDCO™plus-derived cardiac output and arterial-central venous oxygen content difference were compared to 85 simultaneous measurements by indirect calorimetry from study III. Intraclass correlation, Bland-Altman and mixed models analyses for relative changes over time indicated systematic underestimation and poor absolute agreement by this method compared to indirect calorimetry. It may be possible to construct methods for estimation or trending of VO2 from routine monitoring, but further adjustments and assessment in larger populations are needed. In summary, these studies characterise and demonstrate that peri-anaesthetic cardiac output and oxygen consumption undergo changes in elderly patients important to haemodynamic stability and oxygen transport. Mechanistic approaches, with feasible and reliable methods for monitoring or estimation of these changes, are suggested when investigating haemodynamic interventions in elderly

Mechanisms underlying perioperative myocardial injury

Troponin is a biomarker for myocardial injury: it is associated with poor outcomes after noncardiac surgery, but the underlying mechanisms remain unclear. Circulating microRNAs are small non-coding RNAs which regulate post-translational gene expression and are released or secreted after cardiac injury. I examined whether perioperative myocardial injury and acute coronary syndrome share common microRNA signatures by comparing cardiac injury-specific serum microRNAs between matched patients with and without raised postoperative troponin within three days of surgery. MicroRNA expression was quantified using real-time polymerase chain reaction before and after surgery, blinded to troponin status. Cardiac-specific microRNAs showed increased expression after surgery, independent of troponin rise. Bioinformatic analyses identified pathways associated with cellular stress involving adrenergic signalling in the cardiomyocyte, and proteins regulating cardiomyocyte calcium homeostasis. Because pre-existing vagal dysfunction is associated with troponin rise after noncardiac surgery, I hypothesised that an acquired loss of cardiac vagal protective signalling promotes troponin elevation after noncardiac surgery. Serial cardiac autonomic measurements were made in patients before and after noncardiac surgery. Using 5-minute electrocardiogram recordings, cardiac autonomic modulation was quantified by heart rate (HR) variability and HR recovery after a standardised orthostatic manoeuvre. The primary outcome was myocardial injury (high-sensitivity troponin T≥15ng.L¯¹) within 48h of surgery, and the exposure of interest was cardiac vagal activity (high-frequency power spectrum [HFLn]) and HR recovery from peak HR after an orthostatic manoeuvre. HFLn was reduced after 24 hours after surgery in patients who developed myocardial injury and orthostatic HR recovery was slower in patients with myocardial injury, compared with patients who remained free of myocardial injury. Elevations in perioperative troponin may occur due to a lack of cardioprotective vagal mechanisms against adrenergic stress. The data highlight the need for a more considered approach to the interpretation of raised circulating troponin, which is frequently ascribed to an ischaemic aetiology

Cardiac Arrhythmias

The most intimate mechanisms of cardiac arrhythmias are still quite unknown to scientists. Genetic studies on ionic alterations, the electrocardiographic features of cardiac rhythm and an arsenal of diagnostic tests have done more in the last five years than in all the history of cardiology. Similarly, therapy to prevent or cure such diseases is growing rapidly day by day. In this book the reader will be able to see with brighter light some of these intimate mechanisms of production, as well as cutting-edge therapies to date. Genetic studies, electrophysiological and electrocardiographyc features, ion channel alterations, heart diseases still unknown , and even the relationship between the psychic sphere and the heart have been exposed in this book. It deserves to be read

Cardiovascular risk factors for perioperative myocardial injury

PhDBackground: Myocardial injury affects up to one in three patients undergoing non-cardiac surgery. However, very little is known about the underlying pathophysiology. In the general population, patients with elevated resting heart rate are at increased risk of cardiac events, mortality, heart failure and autonomic dysfunction, while hypertension is a well described risk factor for cardiovascular disease. I hypothesised that common abnormalities of heart rate or blood pressure were associated with myocardial injury after non-cardiac surgery. Methods: This thesis comprises a series of secondary analyses of data from five prospective multi-centre epidemiological studies of surgical patients. The main outcome of interest was myocardial injury, defined using objective measurement of cardiac troponin. I used logistic regression analysis to test for association between exposures and outcomes. Results: In a large international cohort, patients with high preoperative heart rate had increased risk of myocardial injury and patients with very low preoperative heart rate had reduced risk of myocardial injury. Patients with elevated preoperative pulse pressure had increased risk of myocardial injury, independent of existing hypertension or systolic blood pressure. High heart rate, or high or low systolic blood pressure during surgery, was associated with increased risk of myocardial injury. In a separate study, elevated preoperative heart rate was associated with cardiopulmonary and autonomic dysfunction, and reduced left ventricular stroke volume, suggestive of heart failure. Finally, autonomic dysfunction, identified using cardiopulmonary exercise testing, was associated with elevated preoperative heart rate, elevated plasma NT-Pro-BNP (indicative of heart failure) and postoperative myocardial injury. Conclusions: Elevated preoperative heart rate, autonomic dysfunction and subclinical heart failure may be part of a common phenotype associated with perioperative myocardial injury. Further research is needed to characterise the pathological processes responsible for myocardial injury, and to identify potential therapeutic targets.Medical Research Council British Journal of Anaesthesia Clinical Research Training Fellowship (grant number MR/M017974/1)

Cardiovascular autonomic control after spinal cord injury: Comprehensive investigations into classification and care

Over 86,000 Canadians live with the consequences of a spinal cord injury (SCI). Injury to spinal autonomic pathways can lead to profound cardiovascular autonomic dysfunction. Key areas of concern identified by individuals living with SCI relate to continence and cardiovascular dysfunction. Conditions that result from autonomic dysfunction, such as autonomic dysreflexia (sudden extreme hypertension) are of particular concern. This thesis examined the cardiovascular autonomic consequences of SCI and their relationship to bowel care, the most potent stimulus for dysreflexia, and a key factor that negatively impacts quality of life after SCI. To assess cardiovascular autonomic control, first a quantitative marker of autonomic dysfunction following SCI had to be identified. In Aim 1 (Chapter 3), cardiovascular dysfunction during, and beyond, the first year of injury (n=63) was assessed using a novel quantitative non-invasive marker of cardiovascular autonomic control. From here, a randomized double-blind placebo-controlled crossover clinical trial to determine the effect of topical afferent blockade (lidocaine) on dysreflexia severity during bowel care was conducted (n=13). Aim 2 (Chapter 4) provides evidence that, contrary to current clinical guidelines, topical lidocaine prolongs bowel care, worsens dysreflexia, and increases cardiovascular symptoms. Despite bowel care concerns, past research shows that individuals do not change bowel care practices, highlighting knowledge translation gaps concerning evidence-based bowel management strategies. To address this, in Aim 3 (Chapter 5), semi-structured interviews (n=13) were used to examine the barriers and facilitators to changing bowel care. The largest influences on changing bowel care and potentially relevant intervention options were identified. Finally, during dysreflexia profound sympathetic stimulation may increase risk for cardiac arrhythmia. Aim 4 (Chapter 6) evaluated susceptibility to arrhythmia in a rodent-model of SCI, the impact of the sympathomimetic drug dobutamine on arrhythmia risk, and the potential mitigating effect of exercise training. SCI increased susceptibility to cardiac arrhythmia, with dobutamine further increasing susceptibility in high-level SCI. Exercise training ameliorated markers of arrhythmia risk during dobutamine. The research conducted in this thesis uses a translational and patient-orientated approach to bridge the gap between physiological understanding and meaningful improvement in the clinical setting for individuals living with cardiovascular and continence implications of SCI

QT interval variability in body surface ECG: measurement, physiological basis, and clinical value: position statement and consensus guidance endorsed by the European Heart Rhythm Association jointly with the ESCWorking Group on Cardiac Cellular Electrophysiology

This consensus guideline discusses the electrocardiographic phenomenon of beat-to-beat QT interval variability (QTV) on surface electrocardiograms. The text covers measurement principles, physiological basis, and clinical value of QTV. Technical considerations include QT interval measurement and the relation between QTV and heart rate variability. Research frontiers of QTV include understanding of QTV physiology, systematic evaluation of the link between QTV and direct measures of neural activity, modelling of the QTV dependence on the variability of other physiological variables, distinction between QTV and general T wave shape variability, and assessing of the QTV utility for guiding therapy. Increased QTV appears to be a risk marker of arrhythmic and cardiovascular death. It remains to be established whether it can guide therapy alone or in combination with other risk factors. QT interval variability has a possible role in non-invasive assessment of tonic sympathetic activity