Mathieu Nadeau

The Kruppel-like transcription factor KLF13 is a novel regulator of heart development.

In humans, congenital heart defects occur in 1–2% of live birth, but the molecular mechanisms and causative genes remain unidentified in the majority of cases. We have uncovered a novel transcription pathway important for heart morphogenesis. We report that KLF13, a member of the Krüppel‐like family of zinc‐finger proteins, is expressed predominantly in the heart, binds evolutionarily conserved regulatory elements on cardiac promoters and activates cardiac transcription. KLF13 is conserved across species and knockdown of KLF13 in Xenopus embryos leads to atrial septal defects and hypotrabeculation similar to those observed in humans or mice with hypomorphic GATA‐4 alleles. Physical and functional interaction with GATA‐4, a dosage‐sensitive cardiac regulator, provides a mechanistic explanation for KLF13 action in the heart. The data demonstrate that KLF13 is an important component of the transcription network required for heart development and suggest that KLF13 is a GATA‐4 modifier; by analogy to other GATA‐4 collaborators, mutations in KLF13 may be causative for congenital human heart disease.

An endocardial pathway involving Tbx5, Gata4, and Nos3 required for atrial septum formation

In humans, septal defects are among the most prevalent congenital heart diseases, but their cellular and molecular origins are not fully understood. We report that transcription factor Tbx5 is present in a subpopulation of endocardial cells and that its deletion therein results in fully penetrant, dose-dependent atrial septal defects in mice. Increased apoptosis of endocardial cells lacking Tbx5, as well as neighboring TBX5-positive myocardial cells of the atrial septum through activation of endocardial NOS (Nos3), is the underlying mechanism of disease. Compound Tbx5 and Nos3 haploinsufficiency in mice worsens the cardiac phenotype. The data identify a pathway for endocardial cell survival and unravel a cell-autonomous role for Tbx5 therein. The finding that Nos3, a gene regulated by many congenital heart disease risk factors including stress and diabetes, interacts genetically with Tbx5 provides a molecular framework to understand gene–environment interaction in the setting of human birth defects.

Hypothermic Total Liquid Ventilation Is Highly Protective Through Cerebral Hemodynamic Preservation and Sepsis-Like Mitigation After Asphyxial Cardiac Arrest.

Objectives:Total liquid ventilation provides ultrafast and potently neuro- and cardioprotective cooling after shockable cardiac arrest and myocardial infarction in animals. Our goal was to decipher the effect of hypothermic total liquid ventilation on the systemic and cerebral response to asphyxial cardiac arrest using an original pressure- and volume-controlled ventilation strategy in rabbits. Design:Randomized animal study. Setting:Academic research laboratory. Subjects:New Zealand Rabbits. Interventions:Thirty-six rabbits were submitted to 13 minutes of asphyxia, leading to cardiac arrest. After resumption of spontaneous circulation, they underwent either normothermic life support (control group, n = 12) or hypothermia induced by either 30 minutes of total liquid ventilation (total liquid ventilation group, n = 12) or IV cold saline (conventional cooling group, n = 12). Measurements and Main Results:Ultrafast cooling with total liquid ventilation (32°C within 5 min in the esophagus) dramatically attenuated the post–cardiac arrest syndrome regarding survival, neurologic dysfunction, and histologic lesions (brain, heart, kidneys, liver, and lungs). Final survival rate achieved 58% versus 0% and 8% in total liquid ventilation, control, and conventional cooling groups (p < 0.05), respectively. This was accompanied by an early preservation of the blood-brain barrier integrity and cerebral hemodynamics as well as reduction in the immediate reactive oxygen species production in the brain, heart, and kidneys after cardiac arrest. Later on, total liquid ventilation also mitigated the systemic inflammatory response through alteration of monocyte chemoattractant protein-1, interleukin-1&bgr;, and interleukin-8 transcripts levels compared with control. In the conventional cooling group, cooling was achieved more slowly (32°C within 90–120 min in the esophagus), providing none of the above-mentioned systemic or organ protection. Conclusions:Ultrafast cooling by total liquid ventilation limits the post–cardiac arrest syndrome after asphyxial cardiac arrest in rabbits. This protection involves an early limitation in reactive oxidative species production, blood-brain barrier disruption, and delayed preservation against the systemic inflammatory response.

Core Body Temperature Control by Total Liquid Ventilation Using a Virtual Lung Temperature Sensor

In total liquid ventilation (TLV), the lungs are filled with a breathable liquid perfluorocarbon (PFC) while a liquid ventilator ensures proper gas exchange by renewal of a tidal volume of oxygenated and temperature-controlled PFC. Given the rapid changes in core body temperature generated by TLV using the lung has a heat exchanger, it is crucial to have accurate and reliable core body temperature monitoring and control. This study presents the design of a virtual lung temperature sensor to control core temperature. In the first step, the virtual sensor, using expired PFC to estimate lung temperature noninvasively, was validated both in vitro and in vivo. The virtual lung temperature was then used to rapidly and automatically control core temperature. Experimentations were performed using the Inolivent-5.0 liquid ventilator with a feedback controller to modulate inspired PFC temperature thereby controlling lung temperature. The in vivo experimental protocol was conducted on seven newborn lambs instrumented with temperature sensors at the femoral artery, pulmonary artery, oesophagus, right ear drum, and rectum. After stabilization in conventional mechanical ventilation, TLV was initiated with fast hypothermia induction, followed by slow posthypothermic rewarming for 1 h, then by fast rewarming to normothermia and finally a second fast hypothermia induction phase. Results showed that the virtual lung temperature was able to provide an accurate estimation of systemic arterial temperature. Results also demonstrate that TLV can precisely control core body temperature and can be favorably compared to extracorporeal circulation in terms of speed.

Thermal Dynamics in Newborn and Juvenile Models Cooled by Total Liquid Ventilation

Background: Total liquid ventilation (TLV) consists in filling the lungs with a perfluorocarbon (PFC) and using a liquid ventilator to ensure a tidal volume of oxygenated, CO2-free and temperature-controlled PFC. Having a much higher thermal capacity than air, liquid PFCs assume that the filled lungs become an efficient heat exchanger with pulmonary circulation. Objective: The objective of the present study was the development and validation of a parametric lumped thermal model of a subject in TLV. Methods: The lungs were modeled as one compartment in which the control volume varied as a function of the tidal volume. The heat transfer in the body was modeled as seven parallel compartments representing organs and tissues. The thermal model of the lungs and body was validated with two groups of lambs of different ages and weights (newborn and juvenile) undergoing an ultrafast mild therapeutic hypothermia induction by TLV. Results: The model error on all animals yielded a small mean error of -0.1 ± 0.4 °C for the femoral artery and 0.0 ± 0.1 °C for the pulmonary artery. Conclusion: The resulting experimental validation attests that the model provided an accurate estimation of the systemic arterial temperature and the venous return temperature. Significance: This comprehensive thermal model of the lungs and body has the advantage of closely modeling the rapid thermal dynamics in TLV. The model can explain how the time to achieve mild hypothermia between newborn and juvenile lambs remained similar despite of highly different physiological and ventilatory parameters. The strength of the model is its strong relationship with the physiological parameters of the subjects, which suggests its suitability for projection to humans.

A Liquid Ventilator Prototype for Total Liquid Ventilation Preclinical Studies

1.1 Context Mechanical ventilation is a life-saving procedure used for treating acute respiratory distress, when the respiratory system is no longer capable of regulating blood gases via pulmonary gas exchange. While conventional mechanical ventilation (CMV) is often sufficient to transiently replace lung function until recovery, the most severe respiratory distress syndromes must be treated either by non conventional mechanical ventilation such as high frequency ventilation or even non ventilator strategies such as extracorporeal gas exchange (Raoof et al., 2010). Large literature data suggest a radical change in ventilator support by replacing the traditional gas mixture with a breathable liquid. This method, called liquid assisted ventilation, leads to the replacement of the air-liquid interface in the alveoli by a liquidliquid interface. Since the 70s, perfluorocarbon liquids (PFC) have been identified as the best candidates to be used in liquid ventilation due to their high oxygen and carbon dioxide solubility (Wolfson & Shaffer, 2005). In addition, they are biochemically stable and bio-inert molecules, available as medical grade products including for respiratory use. Liquid assisted ventilation can be performed either as partial or total liquid ventilation. During partial liquid ventilation, only a fraction of the lungs are filled with perfluorocarbon liquid and a conventional mechanical gas ventilator ensures lung ventilation. In contrast, during total liquid ventilation (TLV), the lungs are completely filled with perfluorocarbon liquid while a dedicated device, called a liquid ventilator, must be used to periodically renew a liquid tidal volume in the lungs. A large number of preclinical studies involving various animal models of acute respiratory distress syndrome have demonstrated clear benefits from total liquid ventilation as compared to all other tested ventilation strategies, including partial liquid ventilation, conventional and high frequency gas ventilation (Hirschl et al., 1996; Wolfson et al., 2008). Among its several theoretical advantages over CMV, TLV is considered less aggressive for the lungs, due to lower positive inspiratory pressures and lower respiratory rates. This is felt to be beneficial in both pediatric and adult respiratory distress syndromes, where repeated alveolar overdistension during CMV contributes to

Assessing the impacts of total liquid ventilation on left ventricular diastolic function in a model of neonatal respiratory distress syndrome

Background Filling the lung with dense liquid perfluorocarbons during total liquid ventilation (TLV) might compress the myocardium, a plausible explanation for the instability occasionally reported with this technique. Our objective is to assess the impacts of TLV on the cardiovascular system, particularly left ventricular diastolic function, in an ovine model of neonatal respiratory distress syndrome. Method Eight newborns lambs, 3.0 ± 0.4 days (3.2 ± 0.3kg) were used in this crossover experimental study. Animals were intubated, anesthetized and paralyzed. Catheters were inserted in the femoral and pulmonary arteries. A high-fidelity pressure catheter was inserted into the left ventricle. Surfactant deficiency was induced by repeated lung lavages with normal saline. TLV was then conducted for 2 hours using a liquid ventilator prototype. Thoracic echocardiography and cardiac output assessment by thermodilution were performed before and during TLV. Results Left ventricular end diastolic pressure (LVEDP) (9.3 ± 2.1 vs. 9.2 ± 2.4mmHg, p = 0.89) and dimension (1.90 ± 0.09 vs. 1.86 ± 0.12cm, p = 0.72), negative dP/dt (-2589 ± 691 vs. -3115 ± 866mmHg/s, p = 0.50) and cardiac output (436 ± 28 vs. 481 ± 59ml/kg/min, p = 0.26) were not affected by TLV initiation. Left ventricular relaxation time constant (tau) slightly increased from 21.5 ± 3.3 to 24.9 ± 3.7ms (p = 0.03). Mean arterial systemic (48 ± 6 vs. 53 ± 7mmHg, p = 0.38) and pulmonary pressures (31.3 ± 2.5 vs. 30.4 ± 2.3mmHg, p = 0.61) were stable. As expected, the inspiratory phase of liquid cycling exhibited a small but significant effect on most variables (i.e. central venous pressure +2.6 ± 0.5mmHg, p = 0.001; LVEDP +1.18 ± 0.12mmHg, p<0.001). Conclusions TLV was well tolerated in our neonatal lamb model of severe respiratory distress syndrome and had limited impact on left ventricle diastolic function when compared to conventional mechanical ventilation.

Lumped Thermal Model of a Newborn Lamb and a Liquid Ventilator in Total Liquid Ventilation

Total liquid ventilation (TLV) is an emerging and promising mechanical ventilation method in which the lungs are filled with a breathable liquid. Perfluorocarbon (PFC) is the predominant liquid of choice due to its high O2 and CO2 solubility. In TLV, a dedicated liquid ventilator ensures gas exchange by renewing a tidal volume of PFC, which is temperature-controlled, oxygenated and free of CO2. A fundamental difference between TLV and conventional mechanical ventilation relates to the fact that PFCs are approximately 1500 times denser than air. This high density provides PFCs with a large heat capacity, turning the lungs into an efficient heat exchanger with circulating blood. The originality of this study is the development of a lumped thermal model of the body as a heat exchanger coupled to a liquid ventilator. The model was validated with an animal experimentation on a newborn lamb with the Inolivent-5.0 liquid ventilator prototype. TLV was initiated with a fast hypothermia induction, followed successively by a slow posthypothermic rewarming, a fast rewarming and finally a second fast hypothermia induction. Results demonstrate that the model was able to aptly predict, in every phase, the temperature of the lungs, the eardrum, the rectum as well as the various compartments of the liquid ventilator.Copyright

Control of rapid hypothermia induction by total liquid ventilation : Preliminary results

Mild therapeutic hypothermia (MTH) consists in cooling the body temperature of a patient to between 32 and 34°C. This technique helps to preserve tissues and neurological functions in multi-organ failure by preventing ischemic injury. Total liquid ventilation (TLV) ensures gas exchange in the lungs with a liquid, typically perfluorocarbon (PFC). A liquid ventilator is responsible for ensuring cyclic renewal of tidal volume of oxygenated and temperature-controlled PFC. Hence, TLV using the lung as a heat exchanger and PFC as a heat carrier allows ultra fast cooling of the whole body which can help improve outcome after ischemic injuries. The present study was aimed to evaluate the control performance and safety of automated ultrarapid MTH induction by TLV. Experimentation was conducted using the Inolivent-5.0 liquid ventilator equipped with a PFC treatment unit that allows PFC cooling and heating from the flow of energy carrier water inside a double wall installed on an oxygenator. A water circulating bath is used to manage water temperature. A feedback controller was developed to modulate inspired PFC temperature and control body temperature. Such a controller is important since, with MTH induction, heart temperature should not reach 28°C because of a high risk of fibrillation. The in vivo experimental protocol was conducted on a male newborn lamb of 4.7 kg which, after anesthetization, was submitted to conventional gas ventilation and instrumented with temperature sensors at the femoral artery, oesophagus, right ear drum and rectum. After stabilization, TLV was initiated with fast automated MTH induction to 33.5°C until stabilization of all temperatures. MTH could be reached safely in 3 minutes at the femoral artery, in 3.6 minutes at the esophagus, in 7.7 minutes at the eardrum and in 15 minutes at the rectum. All temperatures were stable at 33.5 ± 0.5°C within 15 minutes. The present results reveal that ultra-fast MTH induction by TLV with Inolivent-5.0 is safe for the heart while maintaining esophageal and arterial temperature over 32.6°C.

Optimal control of inspired perfluorocarbon temperature for induction of hypothermia by total liquid ventilation in juvenile lamb model

Mild hypothermia is well known for its therapeutic value in cardio- and neuroprotection. Many recent experimental studies have shown that the swiftness of the cooling offered by total liquid ventilation (TLV) holds great promise in achieving maximal therapeutic effect. TLV is an emerging ventilation technique in which the lungs are filled with breathable liquids, namely perfluorocarbons (PFCs). A liquid ventilator ensures subject ventilation by periodically renewing a volume of oxygenated, CO2-free and temperature-controlled breathable PFC. The substantial difference between breathing air and liquid is related to the fact that PFCs have over 500 times the volumetric thermal capacity of air 100% relative humidity. The PFC-filled lungs thus turn into an efficient heat exchanger with pulmonary circulation. The objective of the present study was to compute a posteriori the optimal inspired PFC temperature for ultrafast induction of mild hypothermia by TLV in a juvenile lamb experimentation using direct optimal control. The continuous time model and the discretized cycle-by-cycle model are presented. The control objectives of the direct optimal control are also presented and the results are compared with experimental data in order to validate the improved control performances. The computed direct optimal control showed that the inspired PFC temperature command can be improved to avoid temperature undershoots without altering the cooling performances.