Oxygen blood transport during experimental sepsis: effect of hypothermia*.

OBJECTIVES: The aim of this study was to highlight the link between induced hypothermia and increased survival duration as observed in the septic model developed by the laboratory. To reach this objective, survival duration and blood oxygen transport capacity were measured at two temperatures-38 °C (induced normothermia) and 34 °C (induced hypothermia)-in septic rats. DESIGN: A prospective, randomized, experimental animal study. SETTING: University laboratory. SUBJECTS: Forty-four male Sprague-Dawley rats (median weight, 232 g; range, 200-303 g). INTERVENTIONS: After anesthesia and obtention of the temperature goal, sepsis was induced by cecal ligation and perforation. MEASUREMENTS AND MAIN RESULTS: Sepsis induction led to death 5 hrs 11 mins ± 0 hr 36 mins after cecal ligation and perforation at 38 °C. At this temperature, significant changes in blood oxygen transport capacity were observed in septic rats; Hill number decreased from 2.36 ± 0.10 (baseline group) to 1.99 ± 0.17 (septic group) (p = .008) and oxygen-hemoglobin affinity decreased and P50 increased from 41.40 ± 2.4 Torr (baseline group) to 51.17 ± 14.07 Torr (septic group). Furthermore, in normothermia, a significant increase of creatinine and albumin plasmatic concentrations was observed 4 hrs after sepsis induction. Survival duration was significantly higher in induced hypothermia (7 hrs 22 mins ± 0 hr 12 mins at 34 °C) compared with induced normothermia. At 34 °C, no significant change in blood oxygen transport capacity was observed. In the same way, exposure to 34 °C induced no change in measured plasmatic parameters except an increase in albumin concentration in septic rats compared with the baseline group. CONCLUSIONS: Sepsis led to a decrease of both oxygen hemoglobin cooperativity and affinity at 38 °C. By contrast, no change in these parameters was observed when sepsis was induced during hypothermia. Taken together, these results could be interpreted in normothermia septic rats as an adaptive mechanism that could enhance the release of oxygen at the tissue level. Hypothermia by slowing down sepsis evolution could increase survival duration.

Glycolytic fluxes in European silver eel, Anguilla anguilla: sex differences and temperature sensitivity.

European silver eels migrate 6000 km to their supposed spawning area in the Sargasso sea. As the eel is fasting, this intense swimming activity is realised only with fat stores, involving mainly red muscle i.e. aerobic metabolism. However, eel migration is performed at depth and thus in cold water, both being known to induce changes in muscle energy metabolism. During migration, white and red muscles can operate together or separately in order to counteract the eventual effects of low temperatures and/or high pressures. We have studied the temperature sensitivity (5, 15, and 25 degrees C) of aerobic and anaerobic metabolism in both sexes. At the same temperature, migrating eels have a higher basal glycolytic flux. Moreover, there are temperature and sex effects: anaerobic glycolysis (JB) is more sensitive to cold water whereas aerobic (JA) is more affected by warm. Males, which are less sensitive to cold water, also have higher aerobic fluxes than females. As depth corresponds to low temperature, the possibility that males migrate more deeply than females is discussed. In an ecophysiological context, it is interesting to suppose that males and female eels migrate at different depths in order to optimize their energy utilization by aerobic and / or anaerobic pathways.

Is the silvering process similar to the effects of pressure acclimatization on yellow eels?

To reproduce eels need to migrate over 6000 km and at pressure but before this migration they metamorphose into silver eels (silvering). The question raised in this study is to determine whether and how the silvering process contributes to pressure resistance. As migration represents a long swimming activity mainly performed with slow muscles, the red muscle energetics of pressure exposed silver eels was studied. By comparing these results to what has already been shown in yellow eels, we point out that the effects of the silvering process on pressure resistance are similar to the effects of pressure acclimatization in yellow eels. As previously shown, success of pressure acclimatization depends on membrane fluidity, but we raise the hypothesis that the high-pressure resistance of silver eels is due to more fluid membranes.

Hydrostatic pressure and cellular respiration: are the values observed post-decompression representative of the reality under pressure?

The goal of this article was to assess whether a pressure change can constitute a bias of interpretation of pressure effects on pressure-acclimatized fishes. This work consisted first in a study of the recompression effects of mitochondrial extracts from eels pressure-acclimatized; and then in a study of red muscle fibre compression/decompression. The first experimental series shows a decrease of mitochondrial performances after recompression when compared with the decompressed group. It is concluded that recompression does not allow to get rid of decompression effects. This is confirmed by the second experimental series which show that a decompression induces a stronger reduction of MO2 than the previous compression.

Pressure resistance of aerobic metabolism in eels from different water environments.

Eels from different locations were tested comparing their energetic capacities to migrate by studying muscle (red and white) aerobic metabolism. As the migratory activity corresponds to a lengthy swimming activity at depth, their pressure resistance was evaluated by considering fish response to compression, mitochondrial respiration measured under pressure (101 ATA) and cytochrome c oxidase after 3 days under pressure. The results show that only fish from two of the sites have the metabolic capacities to cope with the high pressure encountered during migration.

In vitro aerobic and anaerobic muscle capacities in the European eel, Anguilla anguilla: effects of a swimming session.

In order to have a general view of metabolic requirements during swimming, in vitro aerobic and anaerobic fluxes were measured in red and white muscles from silver eels and yellow eels which differ in activity levels and nutritional states. These measurements were performed in control eels and after a 4 day swimming session (70% U(crit) in yellow eels, 80% U(crit) in silver eels). A swimming session significantly increases U(crit) from 12% to 18%, depending on the stage, with a significantly higher in vitro energy cost during the yellow stage at the muscle level. In vitro, the swimming session brings about a gain in anaerobic capacities rather than in aerobic ones. Some in vivo hypotheses are proposed.

Effect of exercise training on respiration and reactive oxygen species metabolism in eel red muscle.

This paper deals with the effects of exercise training on oxygen consumption (MO(2)) and ROS metabolism in the red muscle of trained and untrained female silver eels. Their critical swimming speed (U(crit)) was determined before and after a 4-day training (10h of swimming at 70% of U(crit) and 14 h at 50%, every day). The U(crit) of trained eels increased significantly (by about 7%). The in vitro MO(2) and ROS production by the red fibres were higher (not significant) in trained than in untrained eels, but the ROS production/MO(2) ratio was alike in both groups. The antioxidant-enzyme activities and lipoperoxidation index in trained eels were both lower than those of the untrained ones. These biochemical changes related to the increase in U(crit) suggest that such a training session could maintained or even increased aerobic power of the red muscle without deleterious impact by ROS. These regulations could play a role in the eel's swimming performance efficiency.

Effects of high hydrostatic pressure on the pituitary-gonad axis in the European eel, Anguilla anguilla (L.).

European silver eels are thought to undergo sexual maturation during their oceanic reproductive migration from the European continent to their spawning area in the Sargasso Sea. Tracking data and various anatomical and physiological features suggest that silver eels migrate in deep sea, leading us to hypothesise that high hydrostatic pressure (HP) influences the induction of eel reproduction. We subjected female and male silver eels to 101ATA for 3 and 7 weeks, respectively, in a hyperbaric chamber equipped with a freshwater recirculation system. In comparison with control eels kept at 1 ATA, HP effects were tested against the messenger RNA levels of pituitary gonadotropins (LHbeta, FSHbeta) using quantitative real-time RT-PCR. The effects of HP on gonadal activity were estimated by measuring gonadosomatic index, oocyte diameter and plasma levels of vitellogenin (Vtg) and sex steroids (E(2), 11-KT). At the pituitary level, LHbeta expression tended to increase while FSHbeta expression decreased in both sex, leading to an increase in the LHbeta/FSHbeta ratio. This suggests a differential effect of HP on the expression of the two gonadotropins. In females submitted to HP, we observed a significant increase in oocyte diameter and plasma levels of 11-KT and E(2). A similar trend was observed for 11-KT plasma levels in males. In females, Vtg plasma levels also significantly increased, reflecting the stimulatory effect of sex steroids on hepatic vitellogenesis. Our results suggest that HP plays a specific and positive role in eel reproduction but additional environmental and internal factors are necessary to ensure complete sexual maturation.

Effects of high-pressure acclimatization on silver eel (Anguilla anguilla, L.) slow muscle contraction.

As the spawning migration of the eel is supposed to correspond to a long swimming activity at depth, patterns of slow red muscle contraction have been investigated in European silver eel (Anguilla anguilla L.) exposed for 3 weeks to 10.1 MPa hydrostatic pressure. The results show that pressure-acclimated eels (male and female) show a three-fold decrease in maximum isometric stress of twitch and tetanic contractions while time to peak force, time from peak force to 90% relaxation and ratio of twitch tension to tetanic tension remain unchanged. The observed modifications in slow red muscle mechanical properties do not impede the spawning migration of the eel and are possibly partially compensated by an improvement in the efficiency of oxidative phosphorylation. Effects of changes in membrane fluidity are also discussed.

Effects of mild induced hypothermia during experimental sepsis.

OBJECTIVES: The potential advantages of lowering core temperature during sepsis are to lower energy requirement and to activate various cell-protecting pathways. We experimentally investigated whether postconditioning temperature modifications influence survival duration during experimental sepsis. DESIGN: A prospective, randomized, experimental animal study. SETTING: University laboratory. SUBJECTS: Eighteen male Sprague-Dawley rats (median 326 g, range 310-347 g). INTERVENTIONS: After anesthesia, experimental sepsis was induced by cecal ligation and perforation. The animals were subsequently assigned a core temperature range: normothermia (37 degrees C), hyperthermia (42 degrees C), and mild hypothermia (32 degrees C). Anesthesia and analgesia were continuously maintained until death. MEASUREMENTS AND MAIN RESULTS: Plasma lactate and pyruvate concentrations were measured at sepsis induction (H0), 4 hrs later (H4), and/or at the time of death. A significant increase in lactate concentration was observed at the time of death in the 42 degrees C group (p = .04). Lactate-to-pyruvate ratio increased in the 32 degrees C (at H4) and 42 degrees C (at the time of death) groups (p = .04). A linear correlation between a longer survival duration and a lower assigned core temperature was observed (from 61 +/- 10 mins at 42 degrees C to 289 +/- 17 mins at 37 degrees C and to 533 +/- 69 mins at 32 degrees C; R = .959, p < .0001). CONCLUSIONS: The current results demonstrate that postconditioning hypothermia was associated with increased survival duration during experimental sepsis. Whether the observed benefits on survival duration are due to potential impacts on energy metabolism or to an anti-inflammatory effect of hypothermia requires further investigation.

In vitro effect of hydrostatic pressure exposure on hydroxyl radical production in fish red muscle.

The effects of hydrostatic pressure on reactive oxygen species (ROS) production have been studied in vitro on fish red muscle fibres. In the eel, Anguilla anguilla, previous studies have shown that hydrostatic pressure acclimatization improves oxidative phosphorylation efficiency together with a supposed concomitant decrease in electron leak and ROS production. In order to test the hypothesis of an electron leak decrease under pressure, hydroxyl radical (HO*) production and oxygen consumption were measured on fish red muscle fibres directly exposed to hydrostatic pressure. Experiments were performed under two conditions--atmospheric pressure and hydrostatic pressure (16.1 MPa)--on eel and trout (which exhibit low- and high-pressure sensitivity, respectively). This work has permitted, first, the validation of an indirect HO* measurement (in vitro) on fish red muscle and the documentation of reference values for fish. Second, at atmospheric pressure, results show higher oxygen consumption for trout (+40%) than for eel which is accompanied by higher HO* production (+90%); in addition, both species present a positive relationship between HO* production and oxygen consumption. Hydrostatic pressure exposure reverses this relationship for eel but not for trout. These preliminary results only partially verify the proposed hypothesis and further experiments are needed.

Fish at high pressure: a hundred year history.

Two main periods can be considered in the history of fish metabolism under pressure. The first period (roughly from 1870 to 1970) was mainly descriptive: survival times and behavior were studied and some authors described an increase in oxygen consumption under pressure; later, the counteracting effects of high temperature on pressure were mentioned. The second period (from 1970 onwards) was more integrative and two major ways were explored. The first was to use shallow-water fish, experimentally exposed to hydrostatic pressure, which can induce a metabolic state resembling histotoxic hypoxia. The second way was to use deep-living fish which have, when compared to surface fish, muscle enzymes with higher structural stability, lower activity (in relationship with habitat depth) and kinetics that are less sensitive to pressure increase. Using this approach, it was also shown that muscle composition and function were somewhat different at depth and that deep fish are well adapted to pressure partly by maintaining membrane fluidity (homeoviscous theory). Since about 1990, the two above-mentioned approaches have still been pursued but by fewer researchers. Studies on deep-living fish are mainly concerned with enzyme kinetics whereas shallow water fish are used mainly for cellular energetic studies. Regarding this topic, it has been shown that yellow freshwater eels are able to acclimate to high-pressure effects, by optimizing membrane fluidity and composition (as achieved by deep-living fish), by improving oxidative phosphorylation (increase of P/O ratio) and the glycolytic pathway.

Effects of dichloroacetate and ubiquinone infusions on glycolysis activity and thermal sensitivity during sepsis.

Energy-metabolism disturbances during sepsis are characterized by enhanced glycolytic fluxes and reduced mitochondrial respiration. However, it is not known whether these abnormalities are the result of a specific mitochondrial alteration, decreased pyruvate dehydrogenase (PDH) complex activity, depletion of ubiquinone (CoQ(10); electron donor for the mitochondrial complex III), or all 3. In this study we sought to specify metabolism disturbances in a murine model of sepsis, using either a PDH-activator infusion (dichloroacetate, DCA) or CoQ(10) supplementation. After anesthesia, Sprague-Dawley rats received intravenous saline solution (control; n = 5), DCA (n = 5; 20 mg/100 g), or CoQ(10) (n = 5; 1 mg/100 g), before the induction of sepsis. Increased plasma lactate levels and increased muscle glucose content were observed after 4 hours in the control group. In the DCA group, a decrease in the muscle content of lactate (P <.05) and an increase in muscle glucose content (P <.05) were observed at 4 hours, but no lactatemia variation was noted. In the CoQ(10) group, only increased plasma lactate levels were observed. Increased muscle glycolysis fluxes were observed after 4 hours in the control group, but to a slighter degree in both the DCA and CoQ(10) groups. Only DCA restored a normal temperature sensitivity in the hyperthermia range, but we noted no differences in survival time. In conclusion, only DCA infusion restores normal glycolysis function.

Temperature sensitivity of glycolysis during sepsis.

OBJECTIVE: To investigate the temperature sensitivity of glycolysis during sepsis. DESIGN: A prospective, randomized, controlled animal study. SETTING: The Physiological Department of a University Hospital. SUBJECTS: Ten male Sprague-Dawley rats, weighing 400-500 g. INTERVENTIONS: The rats were assigned to either a septic (n = 5) or a sham-control group (n = 5). After anesthesia (H0), experimental sepsis was induced by a cecal ligation and perforation, and the left lateral gastrocnemius was sampled. Four hours later (H4), a second anesthesia was performed to sample the contralateral muscle. The sham-control group underwent the same procedures, but the cecum was neither ligated nor incised. MEASUREMENTS AND MAIN RESULTS: Glycolytic flux (J(B), the rate at which glycogen can be used in muscle) and the transition time (t99 : the time required for the transition from aerobic to anaerobic metabolism) were measured by using spectrophotometry. The measurements were performed at seven different temperature levels, ranging from 32 to 42 degrees C. For each measured variable, the temperature sensitivity of glycolysis was assessed by computing the Q10 values, which is the variation ratio of the measured variable, attributed to a 10 degrees C temperature increase. In control rats, anesthesia and surgical procedures induced a J(B) increase (7.9 +/- 1.6 at H0 vs. 11.9 +/- 2.1 micromol x min-1 x g(tissue) at H4, p<.05) without any t99 variation. Whatever the group (control or septic), the same temperature variation induced an effect that was approximately three times higher in the hypothermia (<37 degrees C) than in the hyperthermia range (>37 degrees C; p<.05). However, a loss in thermal sensitivity was observed in septic rats in the hyperthermia range (Q10 = 1.2 +/- 0.1 for septic animals vs. 2.3 +/- 0.4 for control animals; p<.05). CONCLUSIONS: This study demonstrates that glycolysis is more sensitive to temperature in the hypothermia range than in the hyperthermia range. The loss in thermal sensitivity at >37 degrees C in septic rats suggests that sepsis may induce a dysregulation of glycolysis. From an energetic point of view, this signifies that hyperthermia may by itself impair energy metabolism without improving energy production and thus must be treated during sepsis.