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Joffrey Zoll,1 Jamal Bouitbir,1 Pascal Sirvent,2 Alexis Klein,3 Antoine Charton,1,4 Liliana Jimenez,3 François R Péronnet,5 Bernard Geny,1 Ruddy Richard61Physiology Department, Faculty of Medicine and EA3072, Université de Strasbourg, Strasbourg, 2Clermont Université, Université Blaise Pascal, EA 3533, Laboratoire des Adaptations Métaboliques à l’Exercice en Conditions Physiologiques et Pathologiques, Clermont-Ferrand, 3Danone Research, Centre Daniel Carasso, Palaiseau, 4Department of Anesthesia and Critical Care and EA3072, Hôpital de Hautepierre, Université de Strasbourg, France; 5Kinesiology Department, Université de Montréal, Montréal, QC, Canada; 6Department of Sport Medicine and Functional Explorations and INRA UMR 1019, Faculty of Medicine, Université d’Auvergne, Clermont-Ferrand, FranceBackground: It has been suggested that oxygen (O2) diffusion could be favored in water enriched in O2 by a new electrolytic process because of O2 trapping in water superstructures (clathrates), which could reduce the local pressure/content relationships for O2 and facilitate O2 diffusion along PO2 gradients.Materials and methods: Mitochondrial respiration was compared in situ in saponin-skinned fibers isolated from the soleus muscles of Wistar rats, in solution enriched in O2 by injection or the electrolytic process 1) at an O2 concentration decreasing from 240 µmol/L to 10 µmol/L (132 mmHg to 5 mmHg), with glutamate–malate or N, N, N', N'-tetramethyl-p-phenylenediamine dihydrochloride (TMPD)–ascorbate (with antimycin A) as substrates; and 2) at increasing adenosine diphosphate (ADP) concentration with glutamate–malate as substrate.Results: As expected, maximal respiration decreased with O2 concentration and, when compared to glutamate–malate, the apparent Km O2 of mitochondria for O2 was significantly lower with TMPD–ascorbate with both waters. However, when compared to the water enriched in O2 by injection, the Km O2 was significantly lower with both electron donors in water enriched in O2 by electrolysis. This was not associated with any increase in the sensitivity of mitochondria to ADP; no significant difference was observed for the Km ADP between the two waters.Conclusion: In this experiment, a higher affinity of the mitochondria for O2 was observed in water enriched in O2 by electrolysis than by injection. This observation is consistent with the hypothesis that O2 diffusion can be facilitated in water enriched in O2 by the electrolytic process.Keywords: saponin-skinned fibers, mitochondrial respiration, glutamate–malate, TMPD–ascorbate, Km O2

Antoine Charton,1 François Péronnet,2 Stephane Doutreleau,3 Evelyne Lonsdorfer,3 Alexis Klein,4 Liliana Jimenez,4 Bernard Geny,3 Pierre Diemunsch,1 Ruddy Richard5 1Department of Anesthesia and Critical Care, and EA 3072, Hôpital de Hautepierre; University of Strasbourg, Strasbourg, France; 2Department of Kinesiology, Université de Montréal, Montreal, QC, Canada; 3CHRU of Strasbourg, Physiology and Functional Explorations Department, New Civil Hospital, Strasbourg, France and University of Strasbourg, Faculty of Medicine, Physiology Department, Strasbourg, France; 4Danone Research, Palaiseau, France; 5Department of Sport Medicine and Functional Explorations, CHU Clermont-Ferrand and INRA UMR 1019, CRNH-Auvergne, Clermont-Ferrand, France Background: Oral administration of oxygenated water has been shown to improve blood oxygenation and could be an alternate way for oxygen (O2) supply. In this experiment, tissue oxygenation was compared in anesthetized pigs receiving a placebo or water enriched in O2 by injection or a new electrolytic process. Methods: Forty-two pigs randomized in three groups received either mineral water as placebo or water enriched in O2 by injection or the electrolytic process (10 mL/kg in the stomach). Hemodynamic parameters, partial pressure of oxygen in the arterial blood (PaO2), skin blood flow, and tissue oxygenation (transcutaneous oxygen pressure, or TcPO2) were monitored during 90 minutes of general anesthesia. Absorption and tissue distribution of the three waters administered were assessed using dilution of deuterium oxide. Results: Mean arterial pressure, heart rate, PaO2, arteriovenous oxygen difference, and water absorption from the gut were not significantly different among the three groups. The deuterium to protium ratio was also similar in the plasma, skin, and muscle at the end of the protocol. Skin blood flow decreased in the three groups. TcPO2 slowly decreased over the last 60 minutes of the experiment in the three groups, but when compared to the control group, the values remained significantly higher in animals that received the water enriched in O2 by electrolysis. Conclusions: In this protocol, water enriched in O2 by electrolysis lessened the decline of peripheral tissue oxygenation. This observation is compatible with the claim that the electrolytic process generates water clathrates which trap O2 and facilitate O2 diffusion along pressure gradients. Potential applications of O2-enriched water include an alternate method of oxygen supply. Keywords: transcutaneous oxygen partial pressure determination, tissue oxygenation, oxygenated water, water clathrate

Cardiopulmonary exercise test (CPET) is the key examination in the follow-up of congenital heart disease (CHD), reflecting aerobic metabolism leading to ATP production by mitochondria. Peripheral blood mononuclear cells (PBMCs) and mitochondrial respiration is related to the severity of acquired heart disease and heart failure. To assess PBMC's mitochondrial respiration in patients with CHD and investigate its correlation with CPET and echocardiographic parameters. Peripheral blood sample withdrawal were obtained from 26 adult patients with CHD: tetralogy of Fallot (TOF, n = 10), transposition of great arteries (TGA, n = 6) and left sided obstructive lesions (LSOLs, n = 10) as part of their routine blood tests. Healthy volunteers (n = 15, matched for sex and age) were also recruited. Blood samples were tested for mitochondrial respiration (using the Oroboros Oxygraph system) and production of reactive oxygen species (ROS). CPET and echocardiography were performed the same day. We aim to understand how the energetic metabolism is linked with the functional status of patients with CHD. Based on previous studies, we expect to observe impairments in mitochondrial respiration and increased oxidative stress proportional to the gravity of the cardiopathy and the alteration of the CPET. Preliminary results show a declining trend of the coupling efficiency (or RCR, ratio of mitochondrial respiration) in oxidative phosphorylation compared to healthy controls, notably in ToF group (P = 0.03) (Fig. 1). Surprisingly we did not observe any significant differences in production of ROS (Fig. 1). Work is in ongoing to establish a strong statistical correlation with clinical data, primarily with the severity of the CHD. Exploring energetic metabolism could be a useful non-invasive complementary tool in evaluating the severity of CHD. Further studies, with larger effectives, may confirm these values as reliable biomarkers for prognosis or support other clinical assessments. Finally, mitochondrial modulation could become a new therapeutic target. [ABSTRACT FROM AUTHOR]

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