Increase In Anaerobic Metabolism Research Articles

Combined Effects of Hypocapnic Hyperventilation and Hypoxia on Exercise Performance and Metabolic Responses During the Wingate Anaerobic Test.

Hypoxia during supramaximal exercise reduces aerobic metabolism with a compensatory increase in anaerobic metabolism without affecting exercise performance. A similar response is elicited by preexercise voluntary hypocapnic hyperventilation, but it remains unclear whether hypocapnic hyperventilation and hypoxia additively reduce aerobic metabolism and increase anaerobic metabolism during supramaximal exercise. To address that issue, 12 healthy subjects (8 males and 4 females) performed the 30-second Wingate anaerobic test (WAnT) after (1)spontaneous breathing in normoxia (control, ∼21% fraction of inspired O2 [FiO2]), (2)voluntary hypocapnic hyperventilation in normoxia (hypocapnia, ∼21% FiO2), (3)spontaneous breathing in hypoxia (hypoxia, ∼11% FiO2), or (4)voluntary hypocapnic hyperventilation in hypoxia (combined, ∼11% FiO2). Mean power output during the 30-second WAnT was similar among the control (561 [133]W), hypocapnia (563 [140]W), hypoxia (558 [131]W), and combined (560 [133]W) trials (P = .778). Oxygen uptake during the 30-second WAnT was lower in the hypocapnia (1523 [318]mL/min), hypoxia (1567 [300]mL/min), and combined (1203 [318]mL/min) trials than in the control (1935 [250]mL/min) trial, and the uptake in the combined trial was lower than in the hypocapnia or hypoxia trial (all P < .001). Oxygen deficit, an index of anaerobic metabolism, was higher in the hypocapnia (38.4 [7.3]mL/kg), hypoxia (37.8 [6.8]mL/kg), and combined (40.7 [6.9]mL/kg) trials than in the control (35.0 [6.8]mL/kg) trial, and the debt was greater in the combined trial than in the hypocapnia or hypoxia trial (all P < .003). Our results suggest that voluntary hypocapnic hyperventilation and hypoxia additively reduce aerobic metabolism and increase anaerobic metabolism without affecting exercise performance during the 30-second WAnT.

P38 mitogen activated protein kinase inhibitor improves platelet in vitro parameters and in vivo survival in a SCID mouse model of transfusion for platelets stored at cold or temperature cycled conditions for 14 days.

Platelets for transfusion are stored at room temperature (20–24°C) up to 7 days but decline in biochemical and morphological parameters during storage and can support bacterial proliferation. This decline is reduced with p38MAPK inhibitor, VX-702. Storage of platelets in the cold (4–6°C) can reduce bacterial proliferation but platelets get activated and have reduced circulation when transfused. Thermocycling (cold storage with brief periodic warm ups) reduces some of the effects of cold storage. We evaluated in vitro properties and in vivo circulation in SCID mouse model of human platelet transfusion of platelets stored in cold or thermocycled for 14 days with and without VX-702. Apheresis platelet units (N = 15) were each aliquoted into five storage bags and stored under different conditions: room temperature; cold temperature; thermocycled temperature; cold temperature with VX-702; thermocycled temperature with VX-702. Platelet in vitro parameters were evaluated at 1, 7 and 14 days. On day 14, platelets were infused into SCID mice to assess their retention in circulation by flow cytometry. VX-702 reduced negative platelet parameters associated with cold and thermocycled storage such as an increase in expression of activation markers CD62, CD63 and of phosphatidylserine (marker of apoptosis measured by Annexin binding) and lowered the rise in lactate (marker of increase in anaerobic metabolism). However, VX-702 did not inhibit agonist-induced platelet aggregation indicating that it does not interfere with platelet hemostatic function. In vivo, VX-702 improved initial recovery and area under the curve in circulation of human platelets infused into a mouse model that has been previously validated against a human platelet infusion clinical trial. In conclusion, inhibition of p38MAPK during 14-days platelet storage in cold or thermocycling conditions improved in vitro platelet parameters and platelet circulation in the mouse model indicating that VX-702 may improve cell physiology and clinical performance of human platelets stored in cold conditions.

Impact of inflammation on brain subcellular energetics in anesthetized rats

BackgroundEmerging data suggests that volatile anesthetic agents may have organ protection properties in the setting of critical illness. The purpose of this study was to better understand the effect of inflammation on cerebral subcellular energetics in animals exposed to two different anesthetic agents—a GABA agonist (propofol) and a volatile agent (isoflurane).ResultsForty-eight Sprague–Dawley rats were anesthetized with isoflurane or propofol. In each group, rats were randomized to celiotomy and closure (sham) or cecal ligation and puncture (inflammation [sepsis model]) for 8 h. Brain tissue oxygen saturation and the oxidation state of cytochrome aa3 were measured. Brain tissue was extracted using the freeze-blow technique. All rats experienced progressive increases in tissue oxygenation and cytochrome aa3 reduction over time. Inflammation had no impact on cytochrome aa3, but isoflurane caused significant cytochrome aa3 reduction. During isoflurane (not propofol) anesthesia, inflammation led to an increase in lactate (+ 0.64 vs. − 0.80 mEq/L, p = 0.0061). There were no differences in ADP:ATP ratios between groups. In the isoflurane (not propofol) group, inflammation increased the expression of hypoxia-inducible factor-1α (62%, p = 0.0012), heme oxygenase-1 (67%, p = 0.0011), and inducible nitric oxide synthase (31%, p = 0.023) in the brain. Animals exposed to inflammation and isoflurane (but not propofol) exhibited increased expression of protein carbonyls (9.2 vs. 7.0 nM/mg protein, p = 0.0050) and S-nitrosylation (49%, p = 0.045) in the brain. RNA sequencing identified an increase in heat shock protein 90 and NF-κβ inhibitor mRNA in the inflammation/isoflurane group.ConclusionsIn the setting of inflammation, rats exposed to isoflurane show increased hypoxia-inducible factor-1α expression despite a lack of hypoxia, increased oxidative stress in the brain, and increased serum lactate, all of which suggest a relative increase in anaerobic metabolism compared to propofol. Differences in oxidative stress as well as heat shock protein 90 and NF-κβ inhibitor may account for the differential expression of cerebral hypoxia-inducible factor-1α during inflammation.

THIAMINE DEFICIENCY IN EXTREMELY PREMATURE NEWBORNS

Introduction. The critical state of extremely premature newborns is accompanied by significant oxidative stress, which leads to an increase in anaerobic metabolism; impaired oxygen utilization in mitochondrial oxidation may occur due to thiamine deficiency.&#x0D; The aim of the study – to learn the thiamine status of extremely premature new-borns in the neonatal period follow-up.&#x0D; Research Methods. The study involved 55 of premature newborns with birth weight less than 1000 g and 35 healthy full-term newborns. The indicated children’s blood tests were made in the first day of life and at the end of the neonatal period. The lactate and magnesium were determined by colorimetric method; the pyruvate content was determined enzymatically; the concentration of vitamin B1 was studied using a fluorimeter.&#x0D; Results and Discussion. In the first days of life, all premature infants have a significantly lower level of thiamine in the blood than the normal one, with the low limit of the norm of mature infants, due to the greatest intensity of adaptive reactions and the redistribution of nutrients in fasting. The deficiency of thiamine of extremely premature infants is accompanied by hypomagnesemia. The defining factor of thiamine deficiency of premature infants is morphofunctional immaturity. Deficiency of thiamine of premature infants is correlated with hyperlactatemia throughout the neonatal period. The metabolic features of an organism of a newborn with an extremely low birth weight reflect hypoxic disturbances (increased lactate and pyruvate level, changes in their ratio), which indicates the activation of glycolysis. The highest level of pyruvate was observed in premature infants in the first day of life and significantly exceeded the reference values with subsequent decrease during the first month of life, but remained beyond the norm, resulting in the development of secondary acidosis and persistent decrease in aerobic metabolism. Thiamine deficiency contributes to the pathogenesis of suppression of the pyruvate-dehydrogenase complex, bypassing pyruvate towards lactate overproduction and the development of metabolic acidosis.&#x0D; Conclusion. Deficiency of thiamine in premature newborns may lead to a deeper critical condition.

Effect of long-term thermal challenge on the Antarctic notothenioid Notothenia rossii.

The thermal stability of the Antarctic Ocean raises questions concerning the metabolic plasticity of Antarctic notothenioids to changes in the environmental temperature. In this study, Notothenia rossii survived 90days at 8°C, and their condition factor level was maintained. However, their hepatosomatic (0.29×) index decreased, indicating a decrease in nutrient storage as a result of changes in the energy demands to support survival. At 8°C, the plasma calcium, magnesium, cholesterol, and triglyceride concentrations decreased, whereas the glucose (1.91×) and albumin (1.26×) concentrations increased. The main energy substrate of the fish changed from lipids to glucose due to a marked increase in lactate dehydrogenase activity, as demonstrated by an increase in anaerobic metabolism. Moreover, malate dehydrogenase activity increased in all tissues, suggesting that fish acclimated at 8°C exhibit enhanced gluconeogenesis. The aerobic demand increased only in the liver due to an increase (2.23×) in citrate synthase activity. Decreases in the activities of superoxide dismutase, catalase, and glutathione-S-transferase to levels that are most likely sufficient at 8°C were observed, establishing a new physiological activity range for antioxidant defense. Our findings indicate that N. rossii has some compensatory mechanisms that enabled its long-term survival at 8°C.

Corrigendum: Hypoxic Induced Decrease in Oxygen Consumption in Cuttlefish (Sepia officinalis) Is Associated with Minor Increases in Mantle Octopine but No Changes in Markers of Protein Turnover

[This corrects the article DOI: 10.3389/fphys.2017.00344.].

Can lactate levels be used as a marker of patent ductus arteriosus in preterm babies?

Serum lactate levels provide information on metabolic capacity at the cellular level. In addition, lactate reflects tissue perfusion and oxygenation status. The aim of this study was to determine the usefulness of high lactate levels as a marker in hemodynamically significant patent ductus arteriosus (hsPDA), which may lead to tissue perfusion defects. Preterm infants with gestational age ≤32weeks and birthweight ≤1500g were included. Lactate levels were determined at postnatal 48-72hours before echocardiographic evaluation. Eligible infants were divided into two groups as infants with and without hsPDA. Cut-off values for lactate were taken as lactate >4mmol/L, identified as a high lactate level. Infants were also divided into two groups according to lactate levels as group I: lactate levels >4mmol/L and group II: lactate levels ≤4mmol/L. Haemodynamic PDA and lactate levels were compared. A total of 119 patients with gestational age ≤32weeks and birthweight ≤1500g were included in the study. Fifty patients had echocardiographic hsPDA and 69 patients had no PDA. Twelve (24%) of the patients with hsPDA and 22 (31.9%) of the non-hsPDA patients had a lactate level of 4mmol/L (P=0.392). There was no correlation between hsPDA presence and lactate levels (P=0.35). High lactate levels are multifactorial and usually indicate impairment of tissue perfusion. There are a number of factors that can lead to impaired tissue perfusion in preterm infants. For the first time in this study, it was shown that lactate levels did not significantly increase in the presence of hemodynamically significant PDA. This may be due to the fact that peripheral tissue perfusion in the presence of hemodynamic PDA does not deteriorate enough to cause an increase in anaerobic metabolism.

Time Course of Metabolic Capacities in Paralarvae of the Common Octopus, Octopus vulgaris, in the First Stages of Life. Searching Biomarkers of Nutritional Imbalance.

The culture of the common octopus (Octopus vulgaris) is promising since the species has a relatively short lifecycle, rapid growth, and high food conversion ratios. However, recent attempts at successful paralarvae culture have failed due to slow growth and high mortality rates. Establishing an optimal nutritional regime for the paralarvae seems to be the impeding step in successful culture methods. Gaining a thorough knowledge of food regulation and assimilation is essential for paralarvae survival and longevity under culture conditions. The aim of this study, then, was to elucidate the characteristic metabolic organization of octopus paralarvae throughout an ontogenic period of 12 days post-hatching, as well as assess the effect of diet enrichment with live prey containing abundant marine phospholipids. Our results showed that throughout the ontogenic period studied, an increase in anaerobic metabolism took place largely due to an increased dependence of paralarvae on exogenous food. Our studies showed that this activity was supported by octopine dehydrogenase activity, with a less significant contribution of lactate dehydrogenase activity. Regarding aerobic metabolism, the use of amino acids was maintained for the duration of the experiment. Our studies also showed a significant increase in the rate of oxidation of fatty acids from 6 days after-hatching. A low, although sustained, capacity for de novo synthesis of glucose from amino acids and glycerol was also observed. Regardless of the composition of the food, glycerol kinase activity significantly increased a few days prior to a massive mortality event. This could be related to a metabolic imbalance in the redox state responsible for the high mortality. Thus, glycerol kinase might be used as an effective nutritional and welfare biomarker. The studies in this report also revealed the important finding that feeding larvae with phospholipid-enriched Artemia improved animal viability and welfare, significantly increasing the rate of survival and growth of paralarvae.

Psychophysiological response in parachute jumps, the effect of experience and type of jump

We aimed to analyse the effect of experience and type of parachute jump on the psychophysiological responses of jumpers. We analysed blood oxygen saturation, heart rate, blood glucose, lactate and creatinkinase, leg strength, isometric hand strength, cortical arousal, specific fine motor skills, self-confidence and cognition, and somatic and state anxiety, before and after four different parachute jumps: a sport parachute jump, a manual tactical parachute jump, tandem pilots, and tandem passengers. Independently of the parachute jump, the psychophysiological responses of experienced paratroopers were not affected by the jumps, except for an increase in anaerobic metabolism. Novice parachute jumpers presented a higher psychophysiological stress response than the experienced jumpers, together with a large anticipatory anxiety response before the jump; however, this decreased after the jump, although the high physiological activation was maintained. This information could be used by civil and military paratroopers' instructors to improve their training programmes.

Hypoxic Induced Decrease in Oxygen Consumption in Cuttlefish (Sepia officinalis) Is Associated with Minor Increases in Mantle Octopine but No Changes in Markers of Protein Turnover.

The common cuttlefish (Sepia officinalis), a dominant species in the north-east Atlantic ocean and Mediterranean Sea, is potentially subject to hypoxic conditions due to eutrophication of coastal waters and intensive aquaculture. Here we initiate studies on the biochemical response to an anticipated level of hypoxia. Cuttlefish challenged for 1 h at an oxygen level of 50% dissolved oxygen saturation showed a decrease in oxygen consumption of 37% associated with an 85% increase in ventilation rate. Octopine levels were increased to a small but significant level in mantle, whereas there was no change in gill or heart. There were no changes in mantle free glucose or glycogen levels. Similarly, the hypoxic period did not result in changes in HSP70 or polyubiquinated protein levels in mantle, gill, or heart. As such, it appears that although there was a decrease in metabolic rate there was only a minor increase in anaerobic metabolism as evidenced by octopine accumulation and no biochemical changes that are hallmarks of alterations in protein trafficking. Experiments with isolated preparations of mantle, gill, and heart revealed that pharmacological inhibition of protein synthesis could decrease oxygen consumption by 32 to 42% or Na+/K+ ATPase activity by 24 to 54% dependent upon tissue type. We propose that the decrease in whole animal oxygen consumption was potentially the result of controlled decreases in the energy demanding processes of both protein synthesis and Na+/K+ ATPase activity.

Chlorine inhalation-induced myocardial depression and failure.

Victims of chlorine (Cl2) inhalation that die demonstrate significant cardiac pathology. However, a gap exists in the understanding of Cl2-induced cardiac dysfunction. This study was performed to characterize cardiac dysfunction occurring after Cl2 exposure in rats at concentrations mimicking accidental human exposures (in the range of 500 or 600 ppm for 30 min). Inhalation of 500 ppm Cl2 for 30 min resulted in increased lactate in the coronary sinus of the rats suggesting an increase in anaerobic metabolism by the heart. There was also an attenuation of myocardial contractile force in an ex vivo (Langendorff technique) retrograde perfused heart preparation. After 20 h of return to room air, Cl2 exposure at 500 ppm was associated with a reduction in systolic and diastolic blood pressure as well echocardiographic/Doppler evidence of significant left ventricular systolic and diastolic dysfunction. Cl2 exposure at 600 ppm (30 min) was associated with biventricular failure (observed at 2 h after exposure) and death. Cardiac mechanical dysfunction persisted despite increasing the inspired oxygen fraction concentration in Cl2-exposed rats (500 ppm) to ameliorate hypoxia that occurs after Cl2 inhalation. Similarly ex vivo cardiac mechanical dysfunction was reproduced by sole exposure to chloramine (a potential circulating Cl2 reactant product). These results suggest an independent and distinctive role of Cl2 (and its reactants) in inducing cardiac toxicity and potentially contributing to mortality.

Sepsis Is Associated With Altered Cerebral Microcirculation and Tissue Hypoxia in Experimental Peritonitis*

Alterations in cerebral microvascular blood flow may develop during sepsis, but the consequences of these abnormalities on tissue oxygenation and metabolism are not well defined. We studied the evolution of microvascular blood flow, brain oxygen tension (PbO2), and metabolism in a clinically relevant animal model of septic shock. Prospective randomized animal study. University hospital research laboratory. Fifteen invasively monitored and mechanically ventilated female sheep. The sheep were randomized to fecal peritonitis (n = 10) or a sham procedure (n = 5), and craniectomies were performed to enable evaluation of cerebral microvascular blood flow, PbO2, and metabolism. The microvascular network of the left frontal cortex was evaluated (at baseline, 6, 12, and 18 hr) using sidestream dark-field videomicroscopy. Using an off-line semiquantitative method, functional capillary density and the proportion of small perfused vessels were calculated. PbO2 was measured hourly by a parenchymal Clark electrode, and cerebral metabolism was assessed by the lactate/pyruvate ratio using brain microdialysis; both these systems were placed in the right frontal cortex. In septic animals, cerebral functional capillary density (from 3.1 ± 0.5 to 1.9 ± 0.4 n/mm, p < 0.001) and proportion of small perfused vessels (from 98% ± 2% to 84% ± 7%, p = 0.004) decreased over the 18-hour study period. Concomitantly, PbO2 decreased (61 ± 5 to 41 ± 7 mm Hg, p < 0.001) and lactate/pyruvate ratio increased (23 ± 5 to 36 ± 19, p < 0.001). At 18 hours, when shock was present, animals with a mean arterial pressure less than 65 mm Hg (n = 6) had similar functional capillary density, proportion of small perfused vessels, and PbO2 values but significantly higher lactate/pyruvate ratio (46 ± 18 vs 20 ± 4, p = 0.009) compared with animals with an mean arterial pressure of 65-70 mm Hg (n = 4). Impaired cerebral microcirculation during sepsis is associated with progressive impairment in PbO2 and brain metabolism. Development of severe hypotension was responsible for a further increase in anaerobic metabolism. These alterations may play an important role in the pathogenesis of brain dysfunction during sepsis.

Effects of temperature acclimation on cardiorespiratory performance of the Antarctic notothenioid Trematomus bernacchii

Notothenioid fishes of the Southern Ocean have evolved under cold and stable temperatures for millions of years. Due to rising temperatures in the Southern Ocean, investigating thermal limits and the capacities for inducing a temperature acclimation response in notothenioids has become of increasing interest. Here, we investigated effects of temperature acclimation on cardiorespiratory responses and cardiac and skeletal muscle energy metabolism in a benthic Antarctic notothenioid, Trematomus bernacchii. We acclimated specimens to −1, 2 and 4.5 °C for 14 days and quantified heart rates and ventilation rates during an acute increase in temperature. Ventilation rates showed an effect of acclimation both at initial steady-state acclimation conditions and during an acute temperature increase, suggesting a partial thermal compensatory response. However, acclimation did not affect heart rates at steady-state acclimation conditions and the temperatures at which onset of cardiac arrhythmia occurred, suggesting lack of inducible thermal tolerance in cardiac performance. Citrate synthase (CS), lactate dehydrogenase (LDH) and 3-hydroxyacyl dehydrogenase activities in skeletal muscle tissues suggested acclimation-induced shifts in metabolic fuel preferences, and a marked increase in LDH activity with acclimation to 4.5 °C showed an increase in anaerobic metabolism. In heart tissue, CS and LDH activities decreased with acclimation to 4.5 °C, suggesting reduced cardiac ATP production. Overall, the data suggest a partial acclimatory response to temperature by T. bernacchii and support the hypothesis that reduced cardiac acclimatory capacity may play a role in limiting the thermal plasticity of T. bernacchii.

Influence of Total Intravenous Anesthesia, Entropy and Laparoscopy on Oxidative Stress

Recent studies have correlated postoperative mortality with anesthetic mortality, especially with the depth of anesthesia and systolic blood pressure (SBP). The aim of this study is to evaluate the effects of the depth of total intravenous anesthesia (TIVA) using remifentanil and propofol, performed with monitoring of response entropy (RE) on blood concentrations of oxidative stress markers (TBARS and glutathione) during laparoscopic operations. Twenty adult patients, ASA I, BMI 20-26 kg.m(-2), aged 20 to 40 years, were randomly distributed into two groups: Group I underwent anesthetic-surgical procedure with RE maintained between 45 and 59, and Group II underwent anesthetic-surgical procedure with RE between 30 and 44. In both groups, the remifentanil and propofol infusion was controlled by the effector site (Es), adjusted to maintain RE desired values (Groups I and II) and always assessing the suppression rate (SR). Patients were evaluated in six periods: M1 (immediately before anesthesia), M2 (before tracheal intubation [TI]), M3 (5-minutes after TI), M4 (immediately before pneumoperitoneum [PPT]), M5 (1-minute after PPT), and M6 (1-hour after the operation). The following parameters were assessed at all times: SBP, DBP, HR, RE, SR, TBARS, and glutathione. We found increases in TBARS and glutathione in M5, both in Group I and Group II (p<0.05), with higher values in Group II, and SR in three patients in Group II, immediately after PPT. Increased markers in Group I (M5) suggests an increase in anaerobic metabolism (AM) in the splanchnic circulation while the highest values seen in Group II (GII > GI in M5, p<0.05%) suggest interference of another factor (deep anesthesia) responsible for the increase in AM, probably as a result of increased autonomic nervous system depression and minor splanchnic self-regulation.

The effects of temperature on the physiological response to low oxygen in Atlantic sturgeon

Atlantic sturgeon (Acipenser oxyrhynchus), which are bottom dwelling and migratory fish, experience environmental hypoxia in their natural environment. Atlantic sturgeon, acclimated to either 5 or 15°C, were subjected to a 1 h severe (<10 mm Hg) hypoxia challenge in order to document their physiological responses. We measured hematological parameters, including O(2) transport (hemoglobin, hematocrit), ionic (chloride, osmolality), and metabolic (glucose, lactate) variables under normoxic conditions (~160 mm Hg), immediately following a 1 h exposure to hypoxic water, and following a further 2 h of recovery from this challenge in normoxic water. In a second experiment, we assessed the opercular beat frequency before, during, and after hypoxic exposure. Hemoglobin concentrations and hematocrit were significantly different between fish held at 5°C vs. 15°C and also significantly different between normoxia prior to hypoxia and following recovery. Plasma lactate concentrations increased following hypoxia at both temperatures, indicative of an increase in anaerobic metabolism. In contrast, a significant increase in plasma glucose concentrations in response to hypoxia only occurred at 5°C, suggesting different fuel demands under different temperatures. Changes in opercular beat frequency (OBF) were dependent on temperature. At 5°C, OBF increased upon exposure to hypoxia, but returned to pre-exposure levels within 35 min for the remainder of the experiment. During hypoxia at 15°C, OBF increased very briefly, but then rapidly (within 20 min) decreased to levels below control values. Following a return to normoxia, OBF quickly increased to control levels. Overall, these findings suggest that Atlantic sturgeons are relatively tolerant to short-term and severe hypoxic stress, and the strategies for hypoxia tolerance may be temperature dependent.

Epidermal growth factor protects the heart against low-flow ischemia-induced injury

The role of ErbB4 and ErbB2 in the heart of adult mammals is well established. The heart also expresses ErbB1 (the epidermal growth factor (EGF) receptor), but this receptor has received less attention. We studied the effect of EGF on the response of isolated mouse heart to low-flow ischemia and reperfusion. Reducing perfusate flow to 10% for 30 min resulted in an increase in anaerobic metabolism and the leakage of lactate dehydrogenase during reperfusion. In addition, left ventricle +dP/dt and developed pressure were depressed (20-25%) during reperfusion. The addition of EGF 5 min before and throughout the ischemic period prevented the increase in anaerobic metabolism and the leakage of intracellular lactate dehydrogenase during reperfusion. EGF improved both +dP/dt and developed pressure during ischemia and prevented the decrease in dP/dt during reperfusion. To determine whether the effect of EGF on cell integrity depends on its effect on contractility, we studied nonbeating isolated myocytes. In these cells, anoxia and reoxygenation reduced cell viability by nearly 25%. EGF prevented such a decrease. Our results indicate that, like ErbB4 and ErbB2, ErbB1 also has an important role in the heart of adult animals.

Nitric oxide increases myocardial efficiency in the hypoxia-tolerant turtle Trachemys scripta

Nitric oxide (NO) may influence cardiac mechanical performance relative to O2 consumption by depressing respiration rate and by affecting the excitation-contraction coupling. Such effects of NO should be particularly important during hypoxia in species such as the hypoxia-tolerant turtle Trachemys scripta. In heart ventricle preparations from this species, the ratio of twitch force to O2 consumption increased by approximately 15% during full oxygenation and by approximately 60% during hypoxia in the presence of added L-arginine [the substrate for nitric oxide synthase (NOS)]. This effect was primarily due to a decrease in O2 consumption and may represent an increase in the twitch force obtained per ATP and/or in the ATP obtained per O2. Lactate production during hypoxia did not differ between preparations treated with either L-arginine or asymmetric dimethylarginine (ADMA), an inhibitor of NOS, suggesting that NO does not elicit a compensatory increase in anaerobic metabolism. ADMA did not reverse the effects of L-arginine on O2 consumption significantly, although pre-treatment with ADMA abolished the effect of L-arginine, consistent with the competitive binding of L-arginine and ADMA to NOS. Histochemical studies using the fluorescent probe 4,5-diaminofluorescein diacetate (DAF-2 DA) revealed NO production in the presence of added L-arginine. In conclusion, NO may augment heart contractility obtained per O2 by deceasing O2 consumption without affecting either lactate production or developed force. This effect was particularly pronounced under O2 deficiency and may therefore contribute towards preserving cardiac function and to the overall excellent hypoxic tolerance of the turtle.

Metformin and the intestine

To the Editor: The antihyperglycaemic effect of metformin is generally attributed to a decrease in hepatic glucose output, with some additional effects that increase peripheral glucose uptake and utilisation [1]. The intestine also makes an important contribution to the glucose-lowering effect of metformin, but this is often overlooked because of a paucity of clinical data [2]. Animal studies indicate that metformin can cause intestinal glucose absorption to be delayed and occur more distally along the tract [3, 4]. However, animal studies have also shown that metformin increases glucose utilisation by the intestine, particularly anaerobic glucose metabolism [5–7], and this contributes to an apparent shortfall in the passage of glucose from the luminal to the serosal side of the intestine [3]. Extra lactate delivered into the portal vein is at least partly converted into glucose, increasing glucose turnover after administration of metformin [5,7,8]. We demonstrate here that metformin increases lactate production in human intestinal mucosa, similar to reports in animal studies. Animal studies have long established that very high concentrations of metformin accumulate in the wall of the intestine [9], which may at least partly explain the increase in anaerobic metabolism. We provide confirmatory evidence herein that similarly high concentrations of metformin accumulate in the human intestinal mucosa. In the present study, eight recently diagnosed, drug-naive, obese type 2 diabetic patients [five men, three women; age 55±3 years (mean±SEM); BMI 33±1 kg/m] attended on three occasions after an overnight fast. Patients gave informed consent, and ethical approval was granted by the research ethics committee. Avenous blood sample was taken for plasma glucose assay (automated glucose oxidase method), and a mucosal biopsy sample (approximately 25 mg) was taken on each occasion from the proximal jejunum, under light midazolam sedation, using an adapted paediatric gastroscope. On the first occasion, tissue was washed, divided into two portions, weighed and then incubated for 2 h at 37°C in 1 ml of Krebs Ringer bicarbonate buffer supplemented with 20 mg/ml serum albumin, 10 mmol/l glucose, 10 mol/l insulin with or without 10 mol/l metformin hydrochloride, and maintained in an atmosphere of 95% O2, 5% CO2 [4, 5]. Lactate production was measured as the concentration of lactate in the medium (manual spectrophotometric method). Metformin therapy was initiated at a dosage of 850 mg once daily and titrated up to twice daily after 2–3 weeks. Two repeat proximal jejunal mucosal biopsies were obtained after 6–8 weeks of therapy. A pre-dose sample was obtained in the morning, 12–16 h after the last metformin tablet (taken the evening before), and a post-dose sample was taken 3 h after the morning tablet. Tissue samples were washed in buffered saline, blotted and weighed, and then metformin was extracted with acetonitrile and assayed by gas chromatography [10]. The results showed that incubation of the jejunal biopsy tissue with 10 mol/l metformin increased lactate producDiabetologia (2008) 51:1552–1553 DOI 10.1007/s00125-008-1053-5

Effects of sequential changes from conventional ventilation to high-frequency oscillatory ventilation at increasing mean airway pressures in an ovine model of combined lung and head injury

The objective of this study was to determine the intracranial, cardiovascular and respiratory changes induced by conversion to high-frequency oscillator ventilation from conventional mechanical ventilation at increasing airway pressures. In this study, 11 anaesthetized sheep had invasive cardiovascular and intracranial monitors placed. Lung injury was induced by saline lavage and head injury was induced by inflation of an intracranial balloon catheter. All animals were sequentially converted from conventional mechanical ventilation to high-frequency oscillator ventilation at target mean airway pressures of 16, 22, 28, 34 and 40 cm H(2)O. The mean airway pressure was achieved by adjusting positive end expiratory pressure while on conventional mechanical ventilation, and continuous distending pressures while on high-frequency oscillator ventilation. Cerebral lactate production, oxygen consumption and venous oximetry were measured and analysed in relation to changes in transcranial Doppler flow velocity. Transcranial Doppler profiles together with other physiological parameters were measured at each airway pressure. Cerebral perfusion pressure was significantly lower during high-frequency oscillator ventilation than during conventional mechanical ventilation (CMV: 45, 34, 22, 6, 9 mmHg vs. HFOV: 33, 20, 19, 5, 5 mmHg at airway pressures mentioned above, P = 0.02). Intracranial pressure and cerebrovascular resistance increased with increasing intrathoracic pressures (P = 0.001). Cerebral metabolic indices demonstrated an initial increase in anaerobic metabolism followed by a decrease in cerebral oxygen consumption progressing to cerebral infarction as intrathoracic pressures were further increased in a stepwise fashion. Arterial PaCO(2) increased significantly after converting from conventional mechanical ventilation to high-frequency oscillator ventilation (P = 0.001). However, no difference was observed between conventional mechanical ventilation and high-frequency oscillator ventilation when intracranial pressure, metabolic and transcranial Doppler indices were compared at equivalent mean airway pressures. The use of high positive end expiratory pressure with conventional mechanical ventilation or high continuous distending pressure with high-frequency oscillator ventilation increased intracranial pressure and adversely affected cerebral metabolic indices in this ovine model. Transcranial Doppler is a useful adjunct to intracranial pressure and intracranial venous saturation monitoring when major changes in ventilation strategy are adopted.

Lizard threat display handicaps endurance.

Honest-signalling theory asserts that threat displays reliably advertise attributes that influence fighting success. Endurance, as measured by treadmill performance, predicts the outcome of agonistic interactions among lizards. If threat displays in lizards function to advertise endurance capacity then variation in threat displays should correlate with endurance. I tested this prediction for the duration of threat posturing in male side-blotched lizards (Uta stansburiana) and examined whether threat displays act as quality handicaps, reliable signals that expend the attribute that is advertised. Individual variation in the duration of threat posturing correlated with endurance, while an experimental reduction of endurance diminished the duration of threat posturing. As expected of a quality handicap, endurance fell below baseline after display production. A restriction of aerobic metabolism can account for this effect. In threat posturing, lateral compression of the thorax may interfere with respiration or with circulation, limiting aerobic metabolism and causing a compensatory increase in anaerobic metabolism, thereby generating lactate and diminishing locomotor capacity. Concentrations of lactate measured after display production were higher than baseline, consistent with the proposed mechanism. By restricting aerobic metabolism, the threat posture can act as a quality handicap, simultaneously advertising and expending the endurance capacity of displaying lizards.